cyclin-d1 and Carcinoma--Ductal--Breast

Cyclin D1 is one of the most commonly overexpressed oncogenes in breast cancer, with 45-50% of primary ductal carcinomas overexpressing this oncoprotein. Targeted deletion of the gene encoding cyclin D1 demonstrates an essential role in normal mammary gland development while transgenic studies provide evidence that cyclin D1 is a weak oncogene in mammary epithelium. In a recent exciting development, Yu et al. demonstrate that cyclin D1-deficient mice are resistant to mammary carcinomas induced by c-neu and v-Ha-ras, but not those induced by c-myc or Wnt-1. These findings define a pivotal role for cyclin D1 in a subset of mammary cancers in mice and imply a functional role for cyclin D1 overexpression in human breast cancer.

Topics: Animals; Carcinoma, Ductal, Breast; Cyclin D1; Disease Models, Animal; Female; Gene Expression Regulation, Neoplastic; Mammary Neoplasms, Animal; Mice; Mice, Transgenic

Topics: Apoptosis; Breast Neoplasms; Carcinoma in Situ; Carcinoma, Ductal, Breast; Cyclin D1; Female; G1 Phase; Humans; Loss of Heterozygosity; Precancerous Conditions; Response Elements; Transcriptional Activation; Transfection

The deregulation of cyclin D1 (BCL-1, PRAD1, CCND1) protein, normally synthesized in the G1 phase of the cell cycle, has been implicated in the pathogenesis of some malignant neoplasms, including invasive mammary carcinomas. We used rabbit polyclonal antibody 19 to detect cyclin D1 in 55 infiltrating ductal carcinomas and compared the findings to six important clinicopathologic parameters and cyclin D1 gene amplification. Nuclear immunoreactivity of variable intensity for cyclin D1 was present in 35% of the neoplasms, whereas immunoreactivity of normal mammary epithelial nuclei was absent. No significant correlations were observed between immunoreactivity and patient age, axillary lymph node status, estrogen receptors, progesterone receptors, histologic grade, or any of its three components. There was a correlation between cyclin D1 immunostaining and tumor size (P = 0.013). Fourteen of 15 tumors 2 cm or less were negative, whereas 7 of 12 neoplasms larger than 4 cm were immunopositive. Fifteen percent of the invasive carcinomas had cyclin D1 gene amplification. Of these eight tumors, six showed cyclin D1 immunoreactivity (P = 0.017). In this study, cyclin D1 was detected immunohistochemically in approximately one-third of infiltrating ductal carcinomas; approximately one-third of these had detectable cyclin D1 gene amplification. These results further implicate cyclin D1 in breast tumorigenesis and are additional evidence for the role of cell cycle regulatory proteins in invasive mammary carcinoma.

Topics: Adult; Aged; Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Cyclins; Gene Amplification; Humans; Immunohistochemistry; Middle Aged; Oncogene Proteins

Topics: Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Lobular; Cyclin D1; Cyclooxygenase 2; Female; Humans; Insulin Receptor Substrate Proteins

Using chip array assays, we identified differentially expressed genes via a comparison between luminal A breast cancer subtype and normal mammary ductal cells from healthy donors. In silico analysis confirmed by western blot and immunohistochemistry revealed that C-JUN and C-FOS transcription factors are activated in luminal A patients as potential upstream regulators of these differentially expressed genes. Using a chip-on-chip assay, we identified potential C-JUN and C-FOS targets. Among these genes, the NRIP1 gene was revealed to be targeted by C-JUN and C-FOS. This was confirmed after identification and validation with transfection assays specific binding of C-JUN and C-FOS at consensus binding sites. NRIP1 is not only upregulated in luminal A patients and cell lines but also regulates breast cancer-related genes, including PR, ESR1 and CCND1. These results were confirmed by NRIP1 siRNA knockdown and chip array assays, thus highlighting the putative role of NRIP1 in PGR, ESR1 and CCND1 transcriptional regulation and suggesting that NRIP1 could play an important role in breast cancer ductal cell initiation.

Topics: Adult; Aged; Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Estrogen Receptor alpha; Female; Gene Expression Regulation, Neoplastic; Humans; Intracellular Signaling Peptides and Proteins; JNK Mitogen-Activated Protein Kinases; MCF-7 Cells; Middle Aged; Nuclear Proteins; Nuclear Receptor Interacting Protein 1; Proto-Oncogene Proteins c-fos; Transcriptome

Breast cancer is one of the main causes of malignancies in females. Many prognostic parameters are verified but they do not give sufficient data about patients' outcome. So, we must search for new prognostic and clinicopathological parameters. This study aimed to evaluate immunohistochemical expression of cyclin D1 and PSA in breast carcinoma and their relation to prognosis. It includes 79 specimens of breast invasive duct carcinoma. Overall survival time was available for all patients. There is a statistically significant association between positive PSA expression and lower tumor stage, nodal stage and tumor grade and between negative PSA expression and presence of metastasis (

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Female; Humans; Middle Aged; Prostate-Specific Antigen; Retrospective Studies

Cyclin D1 which is associated with cell cycle regulation is solidly established as an oncogene with an important pathogenetic role in breast carcinomas.. The aim of this study was to relate the Cyclin D1 protein overexpression with the amplification of its gene CCND1 in Estrogen Receptors (ER) positive breast carcinomas, in order to investigate the prognostic effect of their aberrations in relation to ER status, also to correlate the Cyclin D1 overexpression with other prognostic parameters.. Chromogenic in situ hybridization (CISH) was used to identify CCND1 amplification on formalin-fixed paraffin-embedded invasive ductal carcinoma, in which immunohistochemistry (IHC) had previously been performed in order to evaluate the pathological relevance of Cyclin D1 overexpression in human breast cancer (n = 138).. CCND1 amplification was identified in 17/138 (12.3%) tumors and 78/138 (56.5%) tumors have overexpressed Cyclin D1. A significant correlation was identified between CCND1 amplification and Cyclin D1 overexpression (P < 0.001) and both Cyclin D1 and CCND1 were related with ER expression.. Our results show a significant correlation between Cyclin D1 overexpression and CCND1 amplification. Overexpression of Cyclin D1was observed in high proportion of breast cancer which should be considered for routine diagnosis.

Topics: Adult; Aged; Algeria; Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Female; Gene Amplification; Humans; Immunohistochemistry; In Situ Hybridization, Fluorescence; Middle Aged; Prognosis

Topics: Blotting, Western; Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Proliferation; Cyclin D1; Endopeptidases; Female; Humans; Immunohistochemistry; Neoplasm Proteins; RNA, Messenger; Transfection; Tumor Cells, Cultured; Ubiquitin Thiolesterase

The aims of this study were to detect \ CCND1\ , \ C-MYC\ , and \ FGFR1\ amplification using qPCR, confirmation with \ FISH, and to further assess their clinicopathological relevance.. Thirty-five breast tumor samples were analyzed for amplification of the selected genes using modified SYBR \ Green qPCR. The accuracy of the qPCR was assessed by FISH as a gold-standard method.. CCND1\ , \ C-MYC\ , and \ FGFR1 \ amplifications were observed in 34.28%, 28.57%, and 17.14% of the 35 samples, respectively.\ qPCR results were significantly confirmed by FISH and qPCR and FISH showed excellent correlation (P = 0.000). \ CCND1\ amplification \ with tumor stage (P = 0.044), positive metastatic status (P = 0.042), positive family history (P = 0.042), and \ C-MYC\ status (P = 0.005); \ C-MYC\ amplification with tumor size (P = 0.021), tumor grade (P = 0.018), tumor stage (P = 0.032), and \ FGFR1\ status (P < 0.000); and \ FGFR1\ amplification with tumor size (P = 0.041) and positive \ ER\ status (P = 0.042) were statistically associated.. Our findings revealed that the applied qPCR approach could precisely quantify the relative gene copy number. More \ studies with a larger sample size are suggested to confirm the clinicopathological value of \ CCND1\ , \ C-MYC\ , and \ FGFR1\ amplification.

Topics: Adolescent; Adult; Benzothiazoles; Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Diamines; DNA-Binding Proteins; Female; Gene Amplification; Gene Dosage; Humans; In Situ Hybridization, Fluorescence; Middle Aged; Neoplasm Grading; Neoplasm Staging; Organic Chemicals; Quinolines; Real-Time Polymerase Chain Reaction; Receptor, Fibroblast Growth Factor, Type 1; Reproducibility of Results; Transcription Factors; Young Adult

Breast carcinoma proliferative activity, histological grade and commercial molecular tests are all important in prognostication and treatment. There is a particular need for improved, standardised techniques for subclassification of grade 2 breast cancers into low-risk and high-risk prognostic groups. In this study we investigated whether gene expression profiling of five proliferation genes was feasible using breast cancer tissue in a clinical setting and whether these profiles could enhance pathological assessment.. Expression of five proliferation gene mRNAs; Ki-67, STK 15, CCNB1, CCND1 and MYBL2, was quantified in 27 breast carcinomas and compared with Ki-67 proliferation index (PI) and Nottingham mitotic score.. Expression of Ki-67, STK15 and MYBL2 mRNA showed moderate Spearman's correlation with Ki-67 PI (p<0.01), but CCND1 and CCNB1 showed weak, non-significant correlation. Individual gene expression did not associate with mitotic score but combined mRNA expression correlated with both Ki-67 PI (p=0.018) and mitotic score (p=0.03; 0.007).. This study confirms mRNA analysis in breast carcinoma formalin-fixed, paraffin-embedded samples is feasible and suggests gene expression profiling, using a small set of five proliferation genes, has potential in aiding histological grading or assessment of proliferative activity of breast cancers. To fully evaluate the clinical applicability of this approach, a larger cohort study with long-term follow-up data is required.

Topics: Aurora Kinase A; Biomarkers, Tumor; Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Cycle Proteins; Cyclin B1; Cyclin D1; Feasibility Studies; Female; Formaldehyde; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Humans; Ki-67 Antigen; Mitotic Index; Paraffin Embedding; Prognosis; Tissue Fixation; Trans-Activators

The Cyclin D1 protein has been extensively studied over the last decades, for its various roles in physiological processes, both in normal and cancer cells. Gene amplifications and overexpression of CCND1 are frequently reported in several types of cancers, including breast carcinomas, showing the increasing relevance of Cyclin D1 in tumorigenesis. Little is known about the role of this protein in the metastatic process, and the main objective of this study was to evaluate the importance of the CCND1 as a potential marker of tumor progression in breast carcinomas, in a sample collected in Southern Brazil. We studied 41 samples of formalin-fixed paraffin-embedded tissue sections from invasive ductal breast carcinomas subdivided into metastatic (n = 19) and non-metastatic (n = 22) tumors. Gene expression analysis was performed through Quantitative Real-Time PCR and immunohistochemistry. In spite of the higher expression levels of CCND1 mRNA and protein in tumors when compared with the control samples, no differences were observed between the metastatic and non-metastatic groups, suggesting that, in these samples, the expression of CCND1 has no significant influence on the metastatic process. Further studies must be performed in an attempt to clarify the diagnostic and prognostic value of Cyclin D1 in breast cancers, as well as the mechanisms that trigger its overexpression in tumors.

Topics: Adult; Aged; Brazil; Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Disease Progression; Female; Gene Expression; Gene Expression Regulation, Neoplastic; Humans; Lymphatic Metastasis; Middle Aged; Prognosis

Ductal carcinoma in situ (DCIS) is a non-invasive form of breast cancer. It is often associated with invasive ductal carcinoma (IDC), and is considered to be a non-obligate precursor of IDC. It is not clear to what extent these two forms of cancer share low-risk susceptibility loci, or whether there are differences in the strength of association for shared loci.. To identify genetic polymorphisms that predispose to DCIS, we pooled data from 38 studies comprising 5,067 cases of DCIS, 24,584 cases of IDC and 37,467 controls, all genotyped using the iCOGS chip.. Most (67 %) of the 76 known breast cancer predisposition loci showed an association with DCIS in the same direction as previously reported for invasive breast cancer. Case-only analysis showed no evidence for differences between associations for IDC and DCIS after considering multiple testing. Analysis by estrogen receptor (ER) status confirmed that loci associated with ER positive IDC were also associated with ER positive DCIS. Analysis of DCIS by grade suggested that two independent SNPs at 11q13.3 near CCND1 were specific to low/intermediate grade DCIS (rs75915166, rs554219). These associations with grade remained after adjusting for ER status and were also found in IDC. We found no novel DCIS-specific loci at a genome wide significance level of P < 5.0x10(-8).. In conclusion, this study provides the strongest evidence to date of a shared genetic susceptibility for IDC and DCIS. Studies with larger numbers of DCIS are needed to determine if IDC or DCIS specific loci exist.

Topics: Adult; Aged; Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Intraductal, Noninfiltrating; Cyclin D1; Female; Genetic Association Studies; Genotype; Humans; Ki-67 Antigen; Middle Aged; Neoplasm Proteins; Polymorphism, Single Nucleotide; Receptor, ErbB-2; Receptors, Estrogen; Receptors, Progesterone

The diagnosis of intraductal papillary lesions of the breast on core biopsy remains challenging in pathology, with most patients requiring formal surgical excision for a definitive diagnosis. The aim of this study was to determine whether a representative panel of proliferative cell cycle immunohistochemical markers (cyclin A2, cyclin B1 and cyclin D1) could improve the specificity of pathological diagnosis of these lesions.. A series of 68 surgically excised intraductal papillary lesion cases were retrospectively selected, and immunohistochemistry for cyclin A2, cyclin B1 and cyclin D1 was performed.. Cyclin B1 (OR 1.80, 95% CI 1.01 to 3.2, p=0.046) and cyclin D1 (OR 1.13, 95% CI 1.05 to 1.22, p=0.002) expression was independently associated with a diagnosis of malignancy in papillary lesions, although expression was frequently heterogeneous and only focal. Cyclin A2 expression (OR 0.76, 95% CI 0.41 to 1.4, p=0.38) was not associated with a malignant diagnosis in multivariable logistic regression models. All three cyclins displayed high sensitivity (80%-95%) for a diagnosis of malignancy, although cyclin B1 showed a superior specificity of 72.7% compared with the low specificity of cyclins A2 and D1.. Our study has identified for the first time that the expression of key cell cycle markers differs between benign and malignant papillary breast lesions and identified changes to the mitotic marker, cyclin B1, as particularly significant. However, given the low level and heterogeneous nature of expression of these markers, there remains a significant risk of undersampling in core biopsies and thus they are unlikely to be useful in routine clinical practice.

Topics: Adult; Aged; Biomarkers, Tumor; Biopsy; Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Papillary; Cell Cycle; Cell Proliferation; Cyclin A2; Cyclin B1; Cyclin D1; Female; Humans; Immunohistochemistry; Logistic Models; Middle Aged; Multivariate Analysis; Odds Ratio; Papilloma, Intraductal; Predictive Value of Tests; Prognosis; Retrospective Studies; Risk Factors

Human protein arginine N-methyltransferase 2 (PRMT2, HRMT1L1) is a protein that belongs to the arginine methyltransferase family, and it has diverse roles in transcriptional regulation through different mechanisms depending on its binding partners. In this study, we provide evidences for the negative effect of PRMT2 on breast cancer cell proliferation in vitro and in vivo. Morever, cyclin D1, one of the key modulators of cell cycle, was found to be downregulated by PRMT2, and PRMT2 was further shown to suppress the estrogen receptor α-binding affinity to the activator protein-1 (AP-1) site in cyclin D1 promoter through indirect binding with AP-1 site, resulting in the inhibition of cyclin D1 promoter activity in MCF-7 cells. Furthermore, a positive correlation between the expression of PRMT2 and cyclin D1 was confirmed in the breast cancer tissues by using tissue microarray assay. In addition, PRMT2 was found to show a high absent percentage in breast caner cell nuclei and the nuclear loss ratio of PRMT2 was demonstrated to positively correlate with cyclin D1 expression and the increasing tumor grade of invasive ductal carcinoma. Those results offer an essential insight into the effect of PRMT2 on breast carcinogenesis, and PRMT2 nuclear loss might be an important biological marker for the diagnosis of breast cancer.

Topics: Adult; Aged; Animals; Biomarkers, Tumor; Blotting, Western; Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Line, Tumor; Cell Nucleus; Cyclin D1; Female; Gene Expression Regulation, Neoplastic; Heterografts; Humans; Immunohistochemistry; Intracellular Signaling Peptides and Proteins; Mice; MicroRNAs; Middle Aged; Mutagenesis, Site-Directed; Neoplasm Grading; Oligonucleotide Array Sequence Analysis; Protein-Arginine N-Methyltransferases; Real-Time Polymerase Chain Reaction; Tissue Array Analysis; Transfection

A needle biopsy of a mass in the right breast of a 36-year-old woman revealed invasive ductal carcinoma (IDC), and approximately 20% of cancer cells showed unequivocal membranous staining with the HercepTest. After systemic therapy with trastuzumab and paclitaxel followed by FEC (fluorouracil + epirubicin + cyclophosphamide), a right mastectomy was performed. By histological and immunohistochemical examinations, the resected tumor consisted mainly of E-cadherin-negative invasive lobular carcinoma (ILC), and the rest was ERBB2-positive IDC; thus, the diagnosis was mixed ductal and lobular carcinoma. Multiplex ligation-dependent probe amplification and fluorescence in situ hybridization (FISH) analyses revealed that ILC and IDC shared high-level amplification of CCND1 in homogeneously staining regions (HSR) and that IDC had an additional HSR-type amplicon of ERBB2. These findings strongly indicate that IDC and ILC had a common precursor cell with CCND1 amplification. Review of the biopsy specimen with FISH showed IDC with gene amplifications of CCND1 and ERBB2 as a minor component, IDC without amplification of CCND1 or ERBB2 as a major component, and a minute portion of ILC with CCND1 amplification. We speculate that chemotherapy and trastuzumab caused a marked reduction in IDC; however, ILC with CCND1 amplification was resistant to chemotherapy and grew.

Topics: Adult; Antineoplastic Agents; Biomarkers, Tumor; Biopsy, Needle; Breast; Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Lobular; Cyclin D1; DNA Fingerprinting; DNA, Neoplasm; Female; Humans; In Situ Hybridization, Fluorescence; Mastectomy; Multiplex Polymerase Chain Reaction; Receptor, ErbB-2; Treatment Outcome

Crk-like (CRKL) is an adapter protein that has crucial roles in multiple biological processes, including cell proliferation, adhesion, and migration. However, the expression pattern of CRKL protein and its clinical significance in human breast cancers have not been well characterized. In this study, expression of CRKL was evaluated in 108 human invasive ductal carcinoma (IDC) tissues by immunohistochemistry. CRKL protein was upregulated in the cancer tissues compared with adjacent normal mammary glands. Overexpression of CRKL was found in 40 of 108 (37.03 %) breast cancer samples and correlated with advanced p-tumor-node-metastasis stage (p = 0.002), nodal metastasis (p = 0.0323), and tumor size (p = 0.0075). In addition, overexpression of CRKL in the MDA-MB-435 cell line promoted cell proliferation, and small interfering RNA knockdown of CRKL in the MDA-MB-453 cell line inhibited proliferation. Further analysis of cell cycle-related molecules showed that CRKL induced cyclin D1 and phosphorylated extracellular signal-regulated kinase expression. In conclusion, this study demonstrated that overexpression of CRKL correlated with progression and malignant proliferation of human breast cancers.

Topics: Adaptor Proteins, Signal Transducing; Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Line, Tumor; Cell Proliferation; Cyclin D1; Extracellular Signal-Regulated MAP Kinases; Female; Gene Expression; Gene Expression Regulation, Neoplastic; Humans; Middle Aged; Nuclear Proteins; Phosphorylation; Protein Processing, Post-Translational

Signal copy number (SCN) and signal intensity (SI) of subtelomeres (ST) are investigated in auxiliary lymph node (ALN) and buccal (BUC) cells by fluorescence in situ hybridization. The extracted total cell of 38256 and 2309 was, respectively, analyzed from the benign ALN- and BUC-cells of an affected breast cancer patient. The Periodic model was based on ST behavior including normal-, down-, and upregulated clones with diverse SCN. The arm-p/q ratio based signature, as a subtelomeric array, reflects discordance and concordance of ST-behavior within individual chromosomes as a concept of "Individualization of Cells" rather than "Global Insight of Cells". The Periodic charts could be considered as a reliable and refreshable platform through which the cellular evolution could be patterned and characterized. Signature of ST-profile in the BUC and ALN cells and the nature of diverse SCN and SI as quantitative and qualitative value led to modeling the real personalized perspective of cellular evolution. Protein expression of Ki67, Cyclin D1, and Cyclin E was assayed, as a complementary panel. These targets could be applied as the predictive and preventive markers for an early detection at BUC and ALN levels to plan the required managements in the breast cancer patients.

Topics: Aged; Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Transformation, Neoplastic; Chromosomes, Human; Cyclin D1; Cyclin E; Female; Humans; In Situ Hybridization, Fluorescence; Ki-67 Antigen; Lymph Nodes; Models, Biological; Mouth Mucosa; Telomere

The tumor-associated calcium signal transducer 2 (TACSTD2) gene has been reported to be highly expressed in many types of human epithelial cancers, and is associated with tumor metastasis and poor prognosis. The aims of the present investigation were to analyze the TACSTD2 and Cyclin D1 expression at the mRNA and protein levels and to assess its prognostic significance in invasive ductal breast cancer (IDC). The expressions of TACSTD2 and Cyclin D1 in IDC tissues were consistently higher than those in the tumor-adjacent non-malignant tissues by a one-step real-time polymerase chain reaction and immunohistochemistry (P<0.001 and P=0.023, respectively). The statistical analysis of clinicopathologic characteristics and immunohistochemistry by the χ(2) test showed that the high expression of TACSTD2 in IDC was correlated to histological grade (P=0.023), P53 status (P=0.042), Cyclin D1 status (P<0.001), lymph node metastasis (P<0.001), distant metastasis (P=0.004) and TNM staging (P<0.001). Kaplan-Meier survival and Cox regression analyses were performed to evaluate the prognosis of IDC. These analyses also showed that a high TACSTD2 expression (P=0.003), a high Cyclin D1 expression (P=0.041), and lymph node metastasis (P=0.006) were independent prognosis factors. Collectively, our studies demonstrated that the high expression of TACSTD2 correlates with a poor prognosis in IDC.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Adhesion Molecules; Cyclin D1; Female; Humans; Lymphatic Metastasis; Neoplasm Metastasis; Neoplasm Staging; Prognosis; RNA, Messenger; Up-Regulation

Recently an increased interest on Elk1 protein and its role in breast cancer evolution has been noted. This protein is an element of the Ets family of transcription factors and it has been involved in a number of important cell processes through the activation of different genes, in a number of normal tissues as well as in many malignancies.. One hundred and seventy (n = 170) cases of operable breast cancer (invasive ductal, lobular and mixed type breast carcinomas) were randomly selected and investigated for the expression of pElk-1, Ki-67 and Cyclin D1 using immunohistochemistry. Our findings were correlated with tumors' clinicopathologic data and biologic profile.. Activated Elk1 is positively associated with ER (p-value: 0.018) and also shows a positive association of with Cyclin D1 (p-value: <0.001). No relationship was noted between pElk1 and Ki67 (p-value: 0.213). Luminal A and B Her-2 negative breast cancer subtypes were showing greater pElk-1 immunoreactivity compared to Her-2 and Basal breast cancer subtypes, and also a higher staining intensity. No association of the molecule with other clinicopathologic characteristics (tumor size, stage, histological type or lymph node metastases) or disease adverse events (local recurrence, metastasis or death) was evidenced.. Our findings offer a new perspective for the role of pElk-1 in breast neoplasia suggesting a direct relation of this molecule to tumor biology and a putative target of personalized breast cancer therapies, although its prognostic/discriminant role is not supported.

Topics: Biomarkers, Tumor; Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Lobular; Cyclin D1; ets-Domain Protein Elk-1; Female; Humans; Immunohistochemistry; Ki-67 Antigen; Phosphorylation; Prognosis; Receptor, ErbB-2; Receptors, Estrogen

  To study the clinicopathological and prognostic value of cyclin D1 overexpression in patients with breast carcinoma..   Immunohistochemistry was performed on paraffin-embedded tissue specimens from 290 invasive breast carcinomas to detect the proteins cyclin D1, oestrogen receptor (ER), progesterone receptor (PR), p53, c-erbB2, and topoisomerase IIα (topoIIα). Cyclin D1 staining was quantified using a computerized image analysis method. Cyclin D1 overexpression characterized smaller, ER-positive and PR-positive tumours (P = 0.017, P < 0.0001, and P < 0.0001, respectively), of a lower histological and nuclear grade (P = 0.011 and P < 0.0001, respectively), and with reduced expression of topoIIα (P = 0.001) and p53 (P < 0.001). Cyclin D1 was found to have an independent favourable impact on the overall survival of both the unselected cohort of patients (P = 0.011) and of patients with ER-negative and lymph node-positive tumours (P = 0.034 and P = 0.015, respectively). In triple-negative tumours, cyclin D1 overexpression was found to have independent favourable impacts on both overall and relapse-free survival (P = 0.002 for both)..   This is the first immunohistochemical study to dissociate the advantageous prognostic effect of cyclin D1 overexpression from its association with ER expression, and to provide evidence that cyclin D1 overexpression may be a marker of prolonged survival in patient subgroups with aggressive phenotypes.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Disease-Free Survival; Female; Humans; Image Interpretation, Computer-Assisted; Immunohistochemistry; Middle Aged; Neoplasm Grading; Neoplasm Staging; Phenotype; Prognosis; Proportional Hazards Models; Receptors, Estrogen

Strong evidence implicates cyclin D1 in human breast cancer. Nevertheless, the prognostic value of cyclin D1 overexpression in breast cancer is still controversial. This work aims to assess the predictive value of cyclin D1 immunohistochemical expression in invasive breast carcinomas and evaluate its association with clinicopathological parameters, in addition to hormone receptor status and Her2/neu immunohistochemical expression.. This study was conducted on 71 cases of invasive breast carcinoma selected according to the availability of clinical data and paraffin-embedded tissue specimens. Immunohistochemistry was performed for estrogen receptors (ER); progesterone receptors (PR); Her2/neu and cyclin D1. Cyclin D1 expression was assessed and compared to the patients' age, tumor histology, grade, nodal status, tumor size and ER; PR; Her2/neu immunostaining results.. Cyclin D1 nuclear expression was detected in 35% of invasive breast carcinomas. There were statistically significant associations between the cyclin D1 and younger age, small tumor size, negative nodal status, well differentiated and lobular types of breast cancer and estrogen receptor positivity. Cyclin D1 had no association with progesterone receptor or Her2/neu.. Cyclin D1 immunohistochemical expression associates strongly with the approved favourable prognostic factors in primary breast carcinoma, suggesting a favourable predictive value of cyclin D1.

Topics: Biomarkers, Tumor; Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Lobular; Cyclin D1; Female; Humans; Lymphatic Metastasis; Middle Aged; Neoplasm Grading; Prognosis; Receptor, ErbB-2; Receptors, Estrogen; Receptors, Progesterone

ATM protein kinase plays a critical role in maintaining genome integrity by activating a biochemical chain reaction that in turn leads to cell cycle checkpoint activation and repair of DNA damage. Cyclin D1 acts in regulating the G1 phase of the cell cycle. Experimental and clinical studies suggest them to be involved in transformation and tumour progression. To elucidate the role of ATM and cyclin D1 expression in sporadic breast cancer, we investigated the possible link between their RNA expression levels in ductal carcinoma and normal adjacent versus normal breast tissues measured by Taqman real-time PCR in 119 breast tissues. Results showed that cyclin D1 over-expressed in 51.4% of breast tumours, whereas ATM expression was down regulated in 55% of breast tumours compared to both normal adjacent and normal controls (P ≤ 0.01). Cyclin D1 expression in adjacent normal and normal tissues was not significantly differed, whereas ATM expression in normal adjacent was lower than normal control (P ≤ 0.01). Over-expression of cyclin D1 correlated with ER(+) and/or PR(+) (oestrogen/progesterone receptor) status, whereas it mostly under-expressed in HER2(+) (human epidermal growth factor 2) tumours. ATM under-expression was more observed in triple-negative tumours (ER(-), PR(-) and HER2(-)). Our results indicated that reduced expression of the ATM and aberrant cyclin D1 expressions may contribute to the development and/or malignant progression of breast carcinomas also the latter could be involved in the regulation of hormone sensitivity associated with ER and PR.

Topics: Adult; Aged; Aged, 80 and over; Ataxia Telangiectasia Mutated Proteins; Biomarkers, Tumor; Breast; Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Cycle Proteins; Cyclin D1; DNA-Binding Proteins; Female; Humans; Iran; Middle Aged; Neoplasm Staging; Protein Serine-Threonine Kinases; Real-Time Polymerase Chain Reaction; Tumor Suppressor Proteins

In the female population in Asia, systematic investigation concerning alterations in cancer-related genes in breast carcinoma is rare, and the correlation among oncogene or suppressor gene expression with tumor cell apoptosis, cell cycle regulation and tumor cell autophagy remains to be clarified. In this study, a tissue microarray consisting of 360 individual samples from three different breast tissues was generated. By comparing the expression of the tumor-suppressor genes (BRCA1, BECN1, CCND1, PTEN and UVRAG) in ductal breast cancer and normal breast tissues, respectively, we were able to assign changes in the expression of these mRNAs to specific stages and allocate them to define the roles in the multi‑step process of breast carcinogenesis. Tumor-suppressor genes, such as BRCA1 and BECN1, usually had lower signals in the carcinomatous tissues (10.2 and 6.6%) compared to the normal tissues (31 and 32.6%), while stronger positive dots (positive cells >30%) usually existed in the normal tissues. The patients in the oldest age group had the lowest expression rate. Only BECN1 and CCND1 expression showed a significant association with patient age (p=0.030 and p=0.003). A significant association was observed between BRCA1 and BECN1 expression and tumor size (p=0.028 and p=0.021). BECN1 gene expression was positively correlated with UVRAG and PTEN expression (p=0.006 and p=0.000). CCND1 was negatively correlated with PTEN, BECN1 and BRCA1 expression (p=0.011, p=0.000 and p=0.000). Abnormal expression of BRCA1, BECN1, CCND1, PTEN and UVRAG may play a role in human breast carcinogenesis through dysregulated mRNA expression. Overexpressed CCND1 may shorten the G1 phase of the cell cycle, suppress cell apoptosis and contribute to the formation of invasive ductal carcinoma (IDC).

Topics: Adult; Aged; Apoptosis Regulatory Proteins; Beclin-1; BRCA1 Protein; Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Female; G1 Phase; Humans; Membrane Proteins; Middle Aged; PTEN Phosphohydrolase; RNA, Messenger; Tumor Suppressor Proteins

To investigate markers for predicting breast cancer progression, we performed a candidate gene-based study that assessed expression change of three genes, cyclin D1, β-catenin, and metastasis-associated protein-1 (MTA1), involving in aggressive phenotypes of cancerous cells, namely hyperproliferation, epithelial-mesenchymal transition, and global transcriptional regulation.. Specimens were from 150 enrolled female patients, with invasive ductal carcinoma, followed up for more than 10 years. mRNA expression of cyclin D1, β-catenin, and MTA1 in cancerous and noncancerous cells microdissected from the primary tumor site was determined by quantitative real-time PCR. The relationship between mRNA expression levels of the genes and clinicopathologic features was assessed by statistical analysis. Disease-free and overall survival (DFS and OS) were analyzed by Kaplan-Meier analysis with log-rank test and a multivariate Cox regression model.. Cyclin D1 was shown to be overexpressed in late-stage breast cancer (stage III/IV). Breast cancer with lymph node metastasis (LNM) showed significantly higher frequency of overexpressed cyclin D1, β-catenin, and MTA1 (P < 0.05). Patients carrying greater numbers of overexpressed genes had joint effects on increased risk in tumors of advanced stages (P ( trend ) = 0.03) and LNM (P ( trend ) < 0.01). In the LNM-negative group, patients whose tumors with greater number of cyclin D1, β-catenin, and MTA1 overexpressions were associated with poorer clinical outcomes, with hazard ratio of 14.79 for OS (P = 0.015) and 7.54 for DFS (P = 0.015) using multivariate Cox regression analysis during the 10-year follow-up.. Higher expression of cyclin D1, β-catenin, and MTA1 mRNAs in breast cancers may prove effective in predicting unfavorable outcomes of breast cancer.

Topics: Adult; Aged; Aged, 80 and over; beta Catenin; Biomarkers, Tumor; Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Female; Follow-Up Studies; Histone Deacetylases; Humans; Immunoenzyme Techniques; Laser Capture Microdissection; Middle Aged; Neoplasm Grading; Neoplasm Staging; Prognosis; Real-Time Polymerase Chain Reaction; Repressor Proteins; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Survival Rate; Trans-Activators

Recent molecular data pointed towards the possibility of a stepwise dedifferentiation in a subgroup of invasive breast cancer (BC) cases. It was hypothesized that oestrogen receptor positive (ER+) grade 3 (G3) ductal invasive BCs are the end stage of a dedifferentiation process of luminal BC. A progression of luminal A towards luminal B BCs associated with a 'progression through grade' and an increased cell proliferation seemed the obvious explanation. In order to verify this hypothesis on a morphological and immunohistochemical level, we investigated 865 invasive BC cases. All cases were reviewed for the presence of intratumoural heterogeneity in grade of the invasive cancer and the presence of associated ductal carcinoma in situ (DCIS). With the use of tissue microarrays, the molecular subtype was determined and correlated with clinico-pathological features. In addition, all cases were stained for p21, p27, Ki-67, Cyclin D1, bcl-2, p53, and p16 and the results subjected to a biomathematical dependency analysis. The frequency of ER-positivity decreased with tumour size. The frequency of luminal A BC decreased as well, whereas the number of luminal B BCs remained constant. A gradual increase of the frequency of basal-like, HER2-driven and non-expressor BCs with tumour size was seen. In only 1 out of 865 BC cases, both a G1 and a G3 invasive cancer component was seen within the same BC. In two cases, a ductal invasive G1 carcinoma was associated with a poorly-differentiated DCIS. The frequency of columnar cell lesions was evenly distributed over ER+ and ER- ductal invasive G3 carcinomas. The biomathematical analysis gave striking hints against an obligate progression of BC trough grade. In conclusion, our results show that a morphological recognizable striking 'progression through grade' at least in its extreme form from G1 towards G3 is a very rare event in the natural course of invasive BC, including luminal BC.

Topics: Adult; Aged; Aged, 80 and over; Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Intraductal, Noninfiltrating; Cyclin D1; Female; Humans; Ki-67 Antigen; Middle Aged; Neoplasm Grading; Proto-Oncogene Proteins c-bcl-2; Receptor, ErbB-2; Receptors, Estrogen; Tumor Suppressor Protein p53

The aim of this work is to study the validity of cyclin D1 expression, a cell cycle regulatory protein, on (fine needle aspiration cytology) FNAC samples in patients with breast carcinoma using immunostaining technique.. This is a study done on 70 patients with primary breast carcinoma, presented to Cytology Unit, Pathology Department, National Cancer Institute, Cairo University. They underwent preoperative FNAC and diagnosed as breast carcinoma. The cytologic and tissue section slides were subjected to cyclin D1 immunocytochemical staining. Only the nuclear immunoreactivity for cyclin D1 was considered specific. The rate of concordance, and discordance, and kappa value were calculated. Relation between cytologic expression of cyclin D1 and different clinicopathologic parameters was evaluated.. Cyclin D1 immunocytochemical expression was observed in 53/70 cases (75.7%) in cytologic smears. In histologic sections of the corresponding cases, cyclin D1was detected in 48/70 cases (68.6%). The concordance rate of cyclin D1 expression in the FNA and histologic sections was 87.1% while the discordance rate was 12.9%. Kappa showed a value of 0.65. A statistically significant relation was found between cyclin D1 immunocytochemical expression and hormonal status as well as nuclear grade.. Cyclin D1 immunocytochemical expression can be performed successfully on cytologic samples with a high concordance rate and agreement with histologic results. This can help in determining tumor biology, and plan for patients' treatment. The marker showed a significant relation with hormone receptor status and nuclear grade.

Topics: Biomarkers, Tumor; Biopsy, Fine-Needle; Breast; Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Female; Humans; Immunohistochemistry; Lymphatic Metastasis; Neoplasm Grading

To investigate the correlation between CCND1 amplification and CHK1 deletion in breast cancer, and to explore their role in tumorigenesis and progression, a comparative study of the gene copy number changes of CCND1 and CHK1 was performed with dual-colour fluorescence in-situ hybridization (FISH).. Sixty-one infiltrating ductal breast carcinomas with foci of ductal carcinoma in situ (DCIS) components were selected for dual-colour FISH. A strong correlation was found between CCND1 amplification and CHK1 deletion (P<0.0001). Fourteen cases were detected that demonstrated both CCND1 amplification and CHK1 deletion. Interestingly, when comparing the infiltrating and non-invasive areas for the same tumour, we found three cases with CCND1 amplification in the infiltrating areas but not in the DCIS areas. We did not find a CHK1 gene profile difference between infiltrating and DCIS areas in the same lesions.. Our findings suggest that CCND1 amplification and CHK1 deletion are common events in breast cancer, and that the two genetic alterations often coexist. Our data also suggest that CHK1 deletion is an early genetic event in the development of breast cancer and can be detected at the DCIS stage, whereas CCND1 amplification is more likely to be associated with tumour progression.

Topics: Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Intraductal, Noninfiltrating; Cell Transformation, Neoplastic; Checkpoint Kinase 1; Cyclin D1; Disease Progression; Female; Gene Amplification; Gene Deletion; Gene Dosage; Humans; Immunohistochemistry; In Situ Hybridization, Fluorescence; Protein Kinases

Understanding the molecular etiology of cancer and increasing the number of drugs and their targets are critical to cancer management. In our attempt to unravel novel breast-cancer associated proteins, we previously conducted protein expression profiling of the MCF10AT model, which comprises a series of isogenic cell lines that mimic different stages of breast cancer progression. NRD1 expression was found to increase during breast cancer progression. Here, we attempted to confirm the relevance of NRD1 in clinical breast cancer and understand the functional role and mechanism of NRD1 in breast cancer cells. Immunohistochemistry data show that NRD1 expression was elevated in ductal carcinoma in situ and invasive ductal carcinomas compared with normal tissues in 30% of the 26 matched cases studied. Examination of NRD1 expression in tissue microarray comprising >100 carcinomas and subsequent correlation with clinical data revealed that NRD1 expression was significantly associated with tumor size, grade, and nodal status (P < 0.05). Silencing of NRD1 reduced MCF10CA1h and MDA-MD-231 breast-cancer-cell proliferation and growth. Probing the oncogenic EGF signaling pathways revealed that NRD1 knock down did not affect overall downstream tyrosine phosphorylation cascades including AKT and MAPK activation. Instead, silencing of NRD1 resulted in a reduction of overall cyclin D1 expression, a reduction of EGF-induced increase in cyclin D1 expression and an increase in apoptotic cell population compared with control cells.

Topics: Apoptosis; Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Line, Tumor; Cell Proliferation; Cyclin D1; Disease Progression; Epidermal Growth Factor; Female; Gene Expression Regulation, Neoplastic; Gene Silencing; Humans; Immunohistochemistry; Metalloproteases; Mitogen-Activated Protein Kinase Kinases; Phosphorylation; Proto-Oncogene Proteins c-akt; Real-Time Polymerase Chain Reaction; RNA, Small Interfering; Signal Transduction; Tyrosine; Up-Regulation

The transcription factor Pit-1/Pou1f1 regulates GH and prolactin (PRL) secretion in the pituitary gland. Pit-1 expression and GH regulation by Pit-1 have also been demonstrated in mammary gland. However, no data are available on the role of Pit-1 on breast PRL. To evaluate this role, several human breast cancer cell lines were transfected with either the Pit-1 expression vector or a Pit-1 small interference RNA construct, followed by PRL mRNA and protein evaluation. In addition, transient transfection of MCF-7 cells by a reporter construct containing the proximal PRL promoter, and ChIP assays were performed. Our data indicate that Pit-1 regulates mammary PRL at transcriptional level by binding to the proximal PRL promoter. We also found that Pit-1 raises cyclin D1 expression before increasing PRL levels, suggesting a PRL-independent effect of Pit-1 on cell proliferation. By using immunohistochemistry, we found a significant correlation between Pit-1 and PRL expression in 94 human breast invasive ductal carcinomas. Considering the possible role of PRL in breast cancer disorders, the function of Pit-1 in breast should be the focus of further research.

Topics: Animals; Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Division; Cell Line, Tumor; Cyclin D1; Female; Gene Expression Regulation, Neoplastic; Genes, bcl-1; Humans; Mice; Mutagenesis, Site-Directed; Neoplasm Proteins; NIH 3T3 Cells; Prolactin; Promoter Regions, Genetic; RNA Interference; RNA, Small Interfering; Transcription Factor Pit-1; Transcription, Genetic

Multiple different biologically and clinically relevant genes are often amplified in invasive breast cancer, including HER2, ESR1, CCND1, and MYC. So far, little is known about their role in tumor progression. To investigate their significance for tumor invasion, we compared pure ductal carcinoma in situ (DCIS) and DCIS associated with invasive cancer with regard to the amplification of these genes. Fluorescence in situ hybridization (FISH) was performed on a tissue microarray containing samples from 130 pure DCIS and 159 DCIS associated with invasive breast cancer. Of the latter patients, we analyzed the intraductal and invasive components separately. In addition, lymph node metastases of 23 patients with invasive carcinoma were included. Amplification rates of pure DCIS and DCIS associated with invasive cancer did not differ significantly (pure DCIS vs. DCIS associated with invasive cancer: HER2 22.7 vs. 24.2%, ESR1 19.0 vs. 24.1%, CCND1 10.0 vs. 14.8%, MYC 11.8 vs. 6.5%; P > 0.05). Furthermore, we observed a high concordance of the amplification status for all genes if in situ and invasive carcinoma of individual patients were compared. This applied also to the corresponding lymph node metastases. Our results indicate no significant differences between the gene amplification status of DCIS and invasive breast cancer concerning HER2, ESR1, CCND1, and MYC. Therefore, our data suggest an early role of all analyzed gene amplifications in breast cancer development but not in the initiation of invasive tumor growth.

Topics: Adult; Aged; Aged, 80 and over; Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Intraductal, Noninfiltrating; Chi-Square Distribution; Cyclin D1; Estrogen Receptor alpha; Female; Gene Amplification; Gene Expression Regulation, Neoplastic; Genotype; Humans; In Situ Hybridization, Fluorescence; Lymphatic Metastasis; Middle Aged; Neoplasm Invasiveness; Phenotype; Proto-Oncogene Proteins c-myc; Receptor, ErbB-2; Tissue Array Analysis

The epidermal growth factor receptor (EGFR)-family and cyclin-D1 have been extensively studied in breast cancer; however systematic studies that examine protein expression and gene status in the same cohort of patients are lacking. Also emerging evidences suggest existence of a direct EGFR-signaling pathway, which involves cellular transport of EGFR from cell membrane to the nucleus, and transcriptional regulation of the target genes. Thus, we examined the protein expression of membrane EGFR, nuclear EGFR, cyclin-D1 and the corresponding gene status in 113 breast carcinomas by immunohistochemistry and fluorescence in situ hybridization using tissue microarrays. Membrane EGFR overexpression and EGFR gene amplification were detected in 2% cases, while nuclear EGFR was detected in 40% of cases, with 12% having high nuclear EGFR staining. Nuclear EGFR correlated with tumor size (P=0.0005), lymph node metastasis (P=0.0288), Nottingham prognostic index (P=0.0011) and estrogen receptor (ER) expression (P=0.0258) but the letter correlation was observed only in premenopausal group of patients. Strong cyclin-D1 expression and cyclin-D1 gene (CCND1) amplification were found in 64 and 13% of the cases, respectively. Cyclin-D1 expression showed positive correlation with ER (P=0.0113) and inverse correlation with Nottingham prognostic index (P=0.0309) and membrane EGFR (P=0.0201). CCND1 amplification also showed inverse correlation with membrane EGFR (P=0.0420). A strong correlation between membrane EGFR expression and gene amplification (P=0.0035), as well as cyclin-D1 overexpression and gene amplification (P=0.0362), was demonstrated. On univariate analysis cyclin-D1 expression showed a correlation with longer overall survival in the premenopausal group and nuclear EGFR correlated with shorter overall survival in whole cohort as well in the premenopausal group of patients. Multivariate analysis revealed nuclear EGFR to be an independent prognostic factor and showed 3.4 times greater mortality risk for nuclear EGFR+++ patients as compared with nuclear EGFR negative patients (hazard ratio =3.402; P=0.0026).

Topics: Biomarkers, Tumor; Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Nucleus; Cyclin D1; ErbB Receptors; Female; Humans; Immunohistochemistry; In Situ Hybridization, Fluorescence; Neoplasm Staging; Premenopause; Prognosis; Survival Rate; Tissue Array Analysis

Breast cancer associated with BRCA1 and BRCA2 gene mutations differs from non-BRCA tumors in several respects. We determined whether there was any difference in CCND1 (11q13) and ZNF217 (20q13) gene amplification with respect to BRCA status. Of 40 breast cancer samples examined, 15 and 9 were from BRCA1 and BRCA2 mutation carriers, respectively, and 16 from patients without mutation. Fluorescence in situ hybridization showed that eight tumors exhibited CCND1 amplification (20%; 3 BRCA1, 3 BRCA2, 2 non-BRCA). ZNF217 amplification was observed in three of 38 cases (8%; 2 BRCA1, 1 non-BRCA). There was no significant difference in CCND1 and ZNF217 amplification between BRCA1, BRCA2 and non-BRCA tumors. CCND1 amplification was associated with decreased disease-free (P = 0.045) and overall survival (P = 0.015). BRCA1 tumors with CCND1 amplification were estrogen receptor negative, in contrast to CCND1 amplified BRCA2 and non-BRCA tumors, suggesting that concurrent CCND1 amplification and estrogen and progesterone receptor negativity may predict germline BRCA1 gene mutation. All ZNF217 amplified tumors were of the medullary histological type (P = 0.002). There was no statistical correlation between CCND1 and ZNF217 amplification and estrogen receptor, progesterone receptor, and ERBB2 expression and TNM classification. CCND1 amplification did not correlate with EGFR expression.

Topics: Adenocarcinoma, Mucinous; Adult; Apoptosis Regulatory Proteins; BRCA1 Protein; BRCA2 Protein; Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Lobular; Cyclin D1; ErbB Receptors; Female; Gene Amplification; Genetic Predisposition to Disease; Germ-Line Mutation; Humans; Immunoenzyme Techniques; In Situ Hybridization, Fluorescence; Middle Aged; Prognosis; Receptor, ErbB-2; Receptors, Estrogen; Receptors, Progesterone; Trans-Activators; Young Adult

To compare the morphologic and immunohistochemical properties of breast carcinomas from Chinese and Australian women in order to define possible biologic differences between these carcinomas.. Three hundred cases of breast carcinomas were assessed for histologic and immunophenotypic characteristics from the pathology archives of the Changhai and St. Vincent's Hospitals.. The Chinese women had proportionally more grade 2 and 3 tumors, whereas Australian women had a higher proportion of grade 1 tumor. There was a higher proportion of younger patients with a larger tumor and patients with lymph node involvement in the Chinese group as compared with Australian women. There was no difference in rate of estrogen receptor positive tumors between the 2 groups. p53 Expression was statistically more common with less cyclin D1 expression in Chinese as compared with Australian women.. This study indicates that both inherent tumor biology and stage at presentation influence adversely affect the outcome of breast carcinoma in Chinese compared as with Australian women.

Topics: Asian People; Australia; Biomarkers, Tumor; Breast Neoplasms; Carcinoma, Ductal, Breast; China; Cyclin D1; Female; Humans; Immunohistochemistry; Middle Aged; Tumor Suppressor Protein p53; White People

Obesity, a condition characterized by increased fat content and altered secretion of adipokines, is a risk factor for postmenopausal breast cancer. Visfatin has recently been established as a novel adipokine that is highly enriched in visceral fat. Here we report that visfatin regulated proliferation of MCF-7 human breast cancer cells. Exogenous administration of recombinant visfatin increased cell proliferation and DNA synthesis rate in MCF-7 cells. Furthermore, visfatin activated G1-S phase cell cycle progression by upregulation of cyclin D1 and cdk2 expression. Visfatin also increased the expression of matrix metalloproteinases 2, matrix metalloproteinases 9, and vascular endothelial growth factor genes, suggesting that it may function in metastasis and angiogenesis of breast cancer. Taken together, these findings suggest that visfatin plays an important role in breast cancer progression.

Topics: Adipocytes; Adipokines; Adipose Tissue, White; Body Mass Index; Bromodeoxyuridine; Carcinoma, Ductal, Breast; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cyclin D1; Cyclin-Dependent Kinase 2; Cyclins; Female; Gene Expression Regulation, Neoplastic; Humans; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Nicotinamide Phosphoribosyltransferase; Recombinant Proteins; Signal Transduction; Up-Regulation; Vascular Endothelial Growth Factor A

To investigate the expression and association of ER, Ki-67 and cyclinD1 in usual ductal hyperplasia(UDH), atypical ductal hyperplasia (ADH) and ductal carcinoma in situ(DCIS) in the breast. The study included 56 cases of pre-cancerous lesions which were surgically excised at Qi Lu Hospital of Shangdong University. Immunohistochemistry was used to determine the expression of ER, Ki-67 and cyclinD1 and double-labelling immunofluorescence technique was used to observe the coexpression of ER and Ki-67. The expression and distribution of ER-positive cells were significantly different in UDH, ADH and DCIS. The ER-positive cells were much more in UDH than in normal TDLUs (terminal duct lobular units). The distribution of ER-positive cells interspersed amid ER-negative cells within UDH. However , the ER positive cells showed marked increases in ADH and low grade nuclear DCIS (P < 0.05), distributing in almost all constituent cells. The expression of ki-67 and cyclinD1 were significantly different between UDH and DCIS (P < 0.05) , and a positive correlation was found between expression of Ki-67 and morphological classification of pre-cancerous lesions (r = 0.3522, P < 0.05) as well as cyclinD1 (r = 0.3901, P < 0.05). Double-labelling immunofluorescence showed that there was no coexpression of ER and Ki-67 in normal breast tissue. The coexpression of the two markers was found in ADH and increased in DCIS. Overexpression of ER, Ki-67 and cyclinD1 significantly accompanies the transition of normal cells and UDH to ADH and DCIS. The coexpression of ER and ki-67 may present the early change in carcinogenesis of breast cancer.

Topics: Breast; Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Intraductal, Noninfiltrating; Cyclin D1; Female; Fluorescent Antibody Technique, Indirect; Humans; Immunoenzyme Techniques; Ki-67 Antigen; Lymphatic Metastasis; Microscopy, Fluorescence; Precancerous Conditions; Prognosis; Receptors, Estrogen

To characterize the molecular genetic profiles of grade 3 invasive ductal carcinomas of no special type using high-resolution microarray-based comparative genomic hybridization (aCGH) and to identify recurrent amplicons harboring putative therapeutic targets associated with luminal, HER-2, and basal-like tumor phenotypes.. Ninety-five grade 3 invasive ductal carcinomas of no special type were classified into luminal, HER-2, and basal-like subgroups using a previously validated immunohistochemical panel. Tumor samples were microdissected and subjected to aCGH using a tiling path 32K BAC array platform. Selected regions of recurrent amplification were validated by means of in situ hybridization. Expression of genes pertaining to selected amplicons was investigated using quantitative real-time PCR and gene silencing was done using previously validated short hairpin RNA constructs.. We show that basal-like and HER-2 tumors are characterized by "sawtooth" and "firestorm" genetic patterns, respectively, whereas luminal cancers were more heterogeneous. Apart from confirming known amplifications associated with basal-like (1q21, 10p, and 12p), luminal (8p12, 11q13, and 11q14), and HER-2 (17q12) cancers, we identified previously unreported recurrent amplifications associated with each molecular subgroup: 19q12 in basal-like, 1q32.1 in luminal, and 14q12 in HER-2 cancers. PPM1D gene amplification (17q23.2) was found in 20% and 8% of HER-2 and luminal cancers, respectively. Silencing of PPM1D by short hairpin RNA resulted in selective loss of viability in tumor cell lines harboring the 17q23.2 amplification.. Our results show the power of aCGH analysis in unraveling the genetic profiles of specific subgroups of cancer and for the identification of novel therapeutic targets.

Topics: Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Line, Tumor; Cyclin D1; Estrogen Receptor alpha; Gene Amplification; Gene Dosage; Gene Expression Profiling; Gene Silencing; Genes, erbB-1; Genes, erbB-2; Humans; Neoplasm Staging; Phosphoprotein Phosphatases; Protein Phosphatase 2C

Pin-1 has been shown to regulate several phases of the cell cycle and is strikingly overexpressed in many human cancers. Vascular endothelial growth factor (VEGF)-C is a potent lymphangiogenic factor produced by tumor and stromal cells. However, little is known about the roles of Pin-1 and VEGF-C in breast carcinoma. p53 protein and cyclin D1 overexpressions have been shown to play a role as prognostic factors in many human cancers. To better understand the roles of Pin-1 and VEGF-C in breast carcinoma, we evaluated the immunohistochemical expression of Pin-1 and VEGF-C in relationship with p53 protein or cyclin D1 overexpression and clinicopathological parameters in 128 mammary infiltrating duct carcinomas. There was a positive expression in 100% of Pin-1, 88% of VEGF-C, 35% of p53 protein, and 66% of cyclin D1 in the breast carcinoma. Correlation of the positive expression of Pin-1 with tumor grade (p<0.01) and lymph node metastasis or cyclin D1 overexpression (p<0.05, respectively) was statistically significant. Significant correlation was observed between VEGF-C and tumor grade, lymph node metastasis or clinical stage (p<0.01, respectively). These results indicate that elevated Pin-1 or VEGF-C expression is more common in infiltrating duct carcinomas with poor prognostic characteristics and is partly associated with an unfavorable outcome. Given the role of cyclin D1 overexpression in oncogenesis of breast, these results suggest that overexpression of Pin-1 and VEGF-C may promote tumor progression and metastasis.

Topics: Adult; Aged; Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Line, Tumor; Cyclin D1; Female; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Lymphatic Metastasis; Middle Aged; NIMA-Interacting Peptidylprolyl Isomerase; Peptidylprolyl Isomerase; Prognosis; Tumor Suppressor Protein p53; Vascular Endothelial Growth Factor C

Pure invasive micropapillary carcinoma (MPC) is a special histological type that accounts for 0.7-3% of all breast cancers. MPC has a distinctive growth pattern and a more aggressive clinical behaviour than invasive ductal carcinomas of no special type (IDC-NSTs). To define the molecular characteristics of MPCs, we profiled a series of 12 MPCs and 24 grade and oestrogen receptor (ER)-matched IDC-NSTs using high-resolution microarray comparative genomic hybridization (aCGH). In addition, we generated a tissue microarray containing a series of 24 MPCs and performed immunohistochemical analysis with ER, PR, Ki-67, HER2, CK5/6, CK14, CK17, EGFR, topoisomerase-IIalpha, cyclin D1, caveolin-1, E-cadherin, and beta-catenin antibodies. In situ hybridization probes were employed to evaluate the prevalence of amplification of HER2, TOP2A, EGFR, CCND1, MYC, ESR1, and FGFR1 genes. aCGH analysis demonstrated that MPCs significantly differed from IDC-NSTs at the genomic level. Gains of 1q, 2q, 4p, 6p, 6q23.2-q27, 7p, 7q, 8p, 8q, 9p, 10p, 11q, 12p, 12q, 16p, 17p, 17q, 19p, 20p, 20q, and 21q, and losses of 1p, 2p, 6q11.1-q16.3, 6q21-q22.1, 9p, 11p, 15q, and 19q were more prevalent in MPCs. High-level gains/amplifications of 8p12-p11, 8q12, 8q13, 8q21, 8q23, 8q24, 17q21, 17q23, and 20q13 were significantly associated with MPCs. A comparison between 24 MPCs and a series of 48 grade and ER-matched IDC-NSTs revealed that high cyclin D1 expression, high proliferation rates, and MYC (8q24) amplification were significantly associated with MPCs. Our results demonstrate that MPCs have distinct histological features and molecular genetic profiles supporting the contention that they constitute a distinct pathological entity.

Topics: Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Disease Progression; Female; Gene Amplification; Gene Expression Profiling; Genetic Markers; Humans; Immunohistochemistry; Immunophenotyping; In Situ Hybridization, Fluorescence; Oligonucleotide Array Sequence Analysis; Oncogenes

Biomarkers, commonly expressed in breast cancer cells, may be correlated with their expression in breast skin of the same subjects.. The expression of biomarkers in specimens from 33 breast tumours and breast skin from the same subject and from 32 normal controls was studied using immunohistochemical techniques.. (1) In normal women, there are significant correlations between the levels of expression of cyclin D1, bcl-2 and p53 in normal breast epithelial cells and breast skin epithelial cells. (2) These patterns of biomarker expression in normal women are similar in breast cancer and breast skin epithelial cells of women with invasive ductal carcinoma (IDC) and ductal carcinoma in situ (DCIS), but are at significantly higher levels in both breast cancer cells and skin from the same subjects. (3) In normal women, human epidermal growth factor receptor 2 (HER-2) is not expressed in either breast epithelial cells or skin epithelial cells. (4) HER-2 is expressed in the breast skin of some subjects with HER-2-positive breast cancer. (5) Positive oestrogen receptor alpha expression occurs significantly more frequently in the breast skin of women with IDC and DCIS than in normal controls.. The influence of localised breast cancer seems to be systemic, and leads to changes in skin and hair.

Topics: Biomarkers, Tumor; Breast; Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Intraductal, Noninfiltrating; Cyclin D1; Estrogen Receptor alpha; Female; Humans; Neoplasm Proteins; Proto-Oncogene Proteins c-bcl-2; Receptor, ErbB-2; Skin; Tumor Suppressor Protein p53

BP1, a novel transcriptional factor, belongs to DLX family of homeobox genes. Recent researches showed that BP1 gene is correlated to genesis of breast cancer, but its correlation to cell cycle control factor has not been reported yet. This study was to observe the expression of BP1 in breast cancer, and to make clear its correlation to Cyclin D1.. The expression of BP1 and Cyclin D1 in 86 specimens of human breast cancer and 20 specimens of normal breast tissue (3 cm away from primary tumor) was detected by reverse transcription-polymerase chain reaction (RT-PCR). BP1 poly antibody was made and was certificated by Western blot. The expression of BP1 and Cyclin D1 in 86 specimens of human breast cancer were detected by immunohistochemistry; their correlation was analyzed.. The positive rate of BP1 mRNA was significanlty higher in breast cancer than in normal breast tissues (69.8% vs. 0, P < 0.001). The positive rate of Cyclin D1 mRNA was 64.0% in breast cancer. BP1 mRNA and Cyclin D1 mRNA were co-expressed in 52 specimens of breast cancer, and simultaneously negative in 23 specimens (P = 0.227); BP1 protein and Cyclin D1 protein were co-expressed in 43 specimens, and simultaneously negative in 31 specimens (P = 0.146).. BP1 gene is highly expressed in breast cancer. There is co-expression of Cyclin D1 and BP1 in breast cancer. BP1 gene may promote the genesis of breast cancer through regulating the expression of Cyclin D1.

Topics: Adult; Aged; Blotting, Western; Breast; Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Medullary; Cyclin D1; Female; Gene Expression Regulation, Neoplastic; Genes, bcl-1; Homeodomain Proteins; Humans; Immunohistochemistry; Middle Aged; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transcription Factors

To clarify MUC1 patterns in invasive ductal breast carcinoma and to relate them to clinicopathological parameters, coexpression of other biological markers and prognosis.. Samples from 243 consecutive patients with primary ductal carcinoma were incorporated into tissue microarrays (TMAs). Slides were stained for MUC1, oestrogen receptor (ER), progesterone receptor (PR), Her2/neu, p53 and cyclin D1. Apical membrane MUC1 expression was associated with smaller tumours (P = 0.001), lower tumour grades (P < 0.001), PR positivity (P = 0.003) and increased overall survival (OS; P = 0.030). Diffuse cytoplasmic MUC1 expression was associated with cyclin D1 positivity (P = 0.009) and increased relapse-free survival (RFS; P = 0.034). Negativity for MUC1 was associated with ER negativity (P = 0.004), PR negativity (P = 0.001) and cyclin D1 negativity (P = 0.009). In stepwise multivariate analysis MUC1 negativity was an independent predictor of both RFS [hazard ratio (HR) 3.5, 95% confidence interval (CI) 1.5, 8.5; P = 0.005] and OS (HR 14.7, 95% CI 4.9, 44.1; P < 0.001).. The expression pattern of MUC1 in invasive ductal breast carcinoma is related to tumour characteristics and clinical outcome. In addition, negative MUC1 expression is an independent risk factor for poor RFS and OS, besides 'classical' prognostic indicators.

Topics: Adult; Aged; Aged, 80 and over; Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Female; Humans; Immunohistochemistry; Kaplan-Meier Estimate; Middle Aged; Mucin-1; Multivariate Analysis; Prognosis; Receptor, ErbB-2; Receptors, Estrogen; Receptors, Progesterone; Tissue Array Analysis; Treatment Outcome; Tumor Suppressor Protein p53

For the female population in Asia, systematic investigation on alterations of cyclin D1 in breast carcinoma is rare, and correlation between cyclin D1 expression with clinicopathological parameters, survival rate, and other prognostic marker associated with cell cycle is unclear.. Expression of cyclin D1 protein, Ki-67, pRb, and p53 was determined by immunohistochemistry in 18 cases of early breast carcinomas and 80 cases of invasive ductal carcinomas. Genetic alteration of cyclin D1 gene and overexpression of cyclin D1 mRNA were detected by Southern blot and RT-PCR, respectively.. Expression of cyclin D1 is negative in usual ductal hyperplasia (UDH) and atypical ductal hyperplasia (ADH). However, in 52.0% (51/98) of all breast carcinomas, positive expression of cyclin D1 was observed. Five-year survival rate of the patients with positive expression of cyclin D1 (52.7%) is significantly lower than the cases with negative expression of cyclin D1 (72.1%). Positive rate of cyclin D1 protein in invasive ductal carcinoma (52.5%) is slightly higher than overexpression rate (40.8%) of cyclin D1 mRNA but significantly higher than amplification rate of cyclin D1 gene (18.4%). Expression of cyclin D1 is correlated with Ki-67 expression, but not correlated with pRb and p53 expression.. Positive expression of cyclin D1 could serve as a poor prognostic marker for Chinese patients with breast carcinoma independent of nodal metastasis and clinical stage. Expression of cyclin D1 protein is affected more directly by overexpression of cyclin D1 mRNA rather than cyclin D1 gene amplification. The cooperation between pRb and p53 with cyclin D1 protein in the carcinogenesis of breast carcinoma is not supported by the results.

Topics: Adult; Age Distribution; Blotting, Southern; Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Intraductal, Noninfiltrating; China; Cyclin D1; Female; Follow-Up Studies; Gene Amplification; Humans; Immunohistochemistry; Ki-67 Antigen; Lymphatic Metastasis; Middle Aged; Neoplasm Staging; Retinoblastoma Protein; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Survival Rate; Time Factors; Tumor Suppressor Protein p53

C-erbB2 and estrogen receptors (ER) are well known for their cell proliferative capacity. Cyclin D1 is a major downstream target of both c-erbB2 and ER. This study was designed to analyze the expression of c-erbB2, cyclin D1 and ER and their prognostic implications in invasive ductal carcinoma of the breast.. The c-erbB2 status was evaluated by fluorescence in situ hybridization and immunohistochemistry (IHC) and cyclin D1 and ER were evaluated by IHC in 333 invasive breast cancer specimens.. The results of FISH and IHC for c-erbB2 showed 86.7% concordance. The overexpression of c-erbB2 was associated with the high expression of cyclin D1 and the negative expression of ER (P < 0.01 for both). The high expression of cyclin D1 was associated with the positive expression of ER (P < 0.01). When the group of patients who overexpressed c-erbB2 were analyzed, the patients with the low expression of cyclin D1 showed a significantly higher mortality than those with the high expression of cyclin D1 (RR = 3.2; 95% CI, 1.6-6.6). When the group of the high cyclin D1 expression was analyzed, the patients with negative expression of ER showed a significantly higher mortality than those with the positive expression of ER (RR = 2.1; 95% CI, 1.1-3.8).. Higher expression of cyclin D1 was associated with better prognosis in patients with c-erbB2 overexpression, and positive expression of ER was associated with better prognosis in patients with high cyclin D1 expression.

Topics: Adult; Aged; Biomarkers, Tumor; Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Down-Regulation; Female; Follow-Up Studies; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; In Situ Hybridization, Fluorescence; Middle Aged; Multivariate Analysis; Predictive Value of Tests; Prognosis; Proportional Hazards Models; Receptor, ErbB-2; Receptors, Estrogen; Survival Analysis; Time Factors; Up-Regulation

The Wnt/beta-catenin signaling cascade is an important signal transduction pathway in human cancers. Overexpression of beta-catenin and its downstream effector, cyclin D1, is implicated in malignant transformation and acquisition of an invasive tumor phenotype. This study aimed to determine the clinical significance of Wnt/beta-catenin canonical pathway components in breast cancer.. Expression of beta-catenin, dishevelled (Dvl) and cyclin D1 was examined in invasive ductal carcinomas (IDCs) of the breast by immunohistochemical analysis.. Of the 98 IDCs analyzed, 30% of tumors displayed both nuclear and cytoplasmic staining of Dvl protein, while 52% showed nuclear localization. Loss of cell surface beta-catenin was observed in 66% of breast carcinomas, whereas nuclear expression was observed in 48% IDCs. Cyclin D1 overexpression was observed in 60% IDCs; 31/59 (53%) of these tumors showed nuclear expression of beta-catenin, suggesting upregulation of the canonical Wnt/beta-catenin pathway. Our study demonstrates a significant association between nuclear localization of Dvl and beta-catenin (p < 0.01, OR = 15.8).. To our knowledge, this is the first study showing an association between nuclear localization of Dvl and beta-catenin in IDCs and suggests the upregulation of Wnt/beta-catenin pathway components, beta-catenin, Dvl and cyclin D1 in IDCs of the breast.

Topics: Adaptor Proteins, Signal Transducing; Adult; Aged; Aged, 80 and over; beta Catenin; Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Dishevelled Proteins; Female; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Lymphatic Metastasis; Middle Aged; Neoplasm Staging; Phosphoproteins; Signal Transduction; Up-Regulation; Wnt Proteins

Breast tumors are usually classified according to their response to estrogens as hormone-dependent or -independent. In this work, we investigated the role of the proinflammatory cytokine TNF-alpha on the estrogen-receptor-positive T47D breast ductal tumor cells. We have found that TNF-alpha exerts a mitogenic effect, inducing cyclin D1 expression and activation of the transcription factor NF-kappaB. Importantly, activation of NF-kappaB was required for estrogen-induced proliferation and cyclin D1 expression. TNF-alpha enhanced the estrogen response by increasing the levels and availability of NF-kappaB. Chromatin immunoprecipitation analysis suggested that the action of estrogens is mediated by a protein complex that contains the activated estrogen receptor, the nuclear receptor coactivator RAC3 and a member of the NF-kappaB family. Finally, our results demonstrate that activation of this transcription factor could be one of the key signals for estrogen-mediated response.

Topics: Animals; Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Proliferation; Chromatin Immunoprecipitation; Cyclin D1; Estrogens; Female; Humans; Mice; NF-kappa B; Receptors, Estrogen; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha

Owing to the gradual modification of breast tissue in postmenopausal women, there can be differential effects on local oestrogen receptor (ER) expression, with potential impingement on the biological behaviour of cancer cells in the ageing. A series of 45 ductal carcinoma (DC) cases were selected in postmenopausal women who were not being treated with HRT. Immunohistochemical analyses were performed for hormone receptors and Ki67 expression. Fluorescence in situ hybridisation (FISH) analysis was carried out to study CCND1 amplification. The selected population was subdivided into three groups by age and was subjected to statistical studies: linear model analysis, estimation of relative incidence (RI), multivariate analysis, and nonparametric tests were performed to investigate whether there were any links between age and molecular variables in DCs. The results show a low rate of proliferation and high ER expression in the oldest age group. In the same group a close correlation was found between high ER expression and CCN in the older age group D1 amplification (P=0.000), as was a more advanced phenotype in terms of tumour size and presence of positive lymph nodes than in the other age groups considered. The results suggest that ductal breast cancer has a favourable molecular prognosis, especially in extreme old age. In particular, there is an inverse correlation between ageing and proliferation rate despite the presence of an accentuated proliferation stimulus (high ER with CCD1 amplifications) in the oldest group relative to the other groups considered.

Topics: Adult; Age Factors; Aged; Aged, 80 and over; Carcinoma, Ductal, Breast; Cell Proliferation; Cyclin D1; Female; Gene Amplification; Humans; Immunohistochemistry; In Situ Hybridization, Fluorescence; Lymphatic Metastasis; Middle Aged; Prognosis; Receptors, Estrogen

In this study, tumour tissue samples of 85 primary breast cancer patients were evaluated for phosphatase and tensin homolog deleted on chromosome ten (PTEN), cyclin D1 and P27/Kip1 expression patterns. The results were correlated with clinicopathological parameters. Loss of PTEN protein expression was present in 32.5% of the cases. Cyclin D1 was overexpressed in 54.2% and P27/Kip1 in 89.3% of the cases. Statistically significant associations were found between PTEN and cyclin D1 expression patterns, and cyclin D1 expression and tumour size.

Topics: Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p27; Female; Gene Expression Regulation, Neoplastic; Humans; PTEN Phosphohydrolase

To explore the potential prognostic value of cyclin D1 in invasive breast cancer and its correlation with basic histopathological parameters, hormonal status (estrogen [ER] and progesterone receptor [PR]), and bcl-2.. Medical records of 48 patients, diagnosed in 1998, from the Central Database of the Institute of Oncology, Clinical Center University of Sarajevo, were analyzed. The mean follow-up was 61 months (range: 4-103 months). Routine histopathological evaluation was performed for 48 formalin-fixed and paraffin-embedded tissue samples. For immunohistochemical staining, we used monoclonal antibodies for ER, PR, bcl-2, and cyclin D1.. Cyclin D1 expression inversely correlated with tumor grade (P=0.010) and tumor size (P=0.023), whereas significant positive association was found with ER (P=0.001) and bcl-2 (P=0.001) expression. Patients with higher cyclin D1 expression had longer both overall survival (P=0.014) and relapse-free survival (P=0.037). Cox regression analysis for overall survival (OS) showed that lymph node status, ER expression, therapy, and cyclin D1 expression were independent prognostic factors. (P range from 0.003 to 0.04).. Expression of cyclin D1 is associated with better disease outcome in breast cancer.

Topics: Adult; Aged; Aged, 80 and over; Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Female; Follow-Up Studies; Humans; Immunohistochemistry; Middle Aged; Prognosis; Proto-Oncogene Proteins c-bcl-2; Receptors, Estrogen

To investigate the expression of cyclin D1, ER, and PR gene proteins and to analyze their relevance to tumor biological characteristics, chemotherapy effects, diseases free survival (DFS) and overall survival (OS) of patients.. Immunohistochemical staining techniques was used to detect the expression of cyclin D1, ER and PR gene protein in 100 samples of breast infiltrating ductal carcinoma patients, all female, aged 49.49 +/- 10.81 (28 approximately 92).. The positive expression rate of gene protein was 60& for ER 58% for PR, and 55% for cyclin D1 ER, PR presented a negative correlation to SBR grading. Patients with cyclin D1 positive tumor had longer OS than those with cyclin D1 negative tumor (P = 0.0053). Coexpression of cyclin D1 and ER was significantly correlated with longer DFS and OS (P(dfs) = 0.0108, P(os) = 0.0030). The positivity of ER and PR was significantly correlated with longer DFS (P(ER) = 0.0322, P(PR) = 0.0129). For those patients receiving postoperative adjuvant chemotherapy, the expression of ER and PR were correlated with a better prognosis and longer DFS. The patients with cyclin D1 negative tumor, who received CAF, had a mean DFS of 51.6 months and a mean OS of 57 months in comparison with 24.8 months and 31.2 months for those patients who received other chemotherapy.. In breast infiltrating ductal carcinoma patients expression of ER and of PR are correlated with longer DFS, cyclin D1 expression is correlated with longer OS. For the patients receiving postoperative adjuvant chemotherapy, the expression of ER and PR correlated to a better prognosis and longer DFS. The patients with cyclin D1 negative tumor who receive CAF chemotherapy have longer DFS and OS than those receiving other chemotherapy.

Topics: Adult; Aged; Aged, 80 and over; Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Female; Humans; Middle Aged; Receptors, Estrogen; Receptors, Progesterone

A(3) adenosine receptor (A(3)AR) activation was shown to inhibit the growth of various tumor cells via the down-regulation of nuclear factor kappaB and cyclin D1. To additionally elucidate whether A(3)AR is a specific target, a survey of its expression in tumor versus adjacent normal cells was conducted.. A(3)AR mRNA expression in various tumor tissues was tested in paraffin-embedded slides using reverse transcription-PCR analysis. A comparison with A(3)AR expression in the relevant adjacent normal tissue or regional lymph node metastasis was performed. In addition, A(3)AR protein expression was studied in fresh tumors and was correlated with that of the adjacent normal tissue.. Reverse transcription-PCR analysis of colon and breast carcinoma tissues showed higher A(3)AR expression in the tumor versus adjacent non-neoplastic tissue or normal tissue. Additional analysis revealed that the lymph node metastasis expressed even more A(3)AR mRNA than the primary tumor tissue. Protein analysis of A(3)AR expression in fresh tumors derived from colon (n = 40) or breast (n = 17) revealed that 61% and 78% had higher A(3)AR expression in the tumor versus normal adjacent tissue, respectively. The high A(3)AR expression level in the tumor tissues was associated with elevated nuclear factor kappaB and cyclin D1 levels. High A(3)AR mRNA expression was also demonstrated in other solid tumor types.. Primary and metastatic tumor tissues highly express A(3)AR indicating that high receptor expression is a characteristic of solid tumors. These findings and our previous data suggest A(3)AR as a potential target for tumor growth inhibition.

Topics: Blotting, Western; Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Line, Tumor; Colonic Neoplasms; Cyclin D1; Down-Regulation; Humans; Lung Neoplasms; Lymphatic Metastasis; Melanoma; Neoplasm Metastasis; Neoplasms; NF-kappa B; Receptor, Adenosine A3; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

E-cadherin links to the cytoskeleton via catenins and mediates cell-cell homophilic adhesion. beta-catenin not only regulates cell-cell adhesion as a protein interacting with cadherin, but also functions as an important component of the Wnt signaling pathway which has been found to be closely associated with tumor formation. This study was performed to examine the expression of E-cadherin, beta-catenin, and cyclin D1 in breast cancer in order to evaluate their possible roles in the formation and progression of breast cancer.. The alterations of E-cadherin, beta-catenin, and cyclin D1 in 60 cases of breast cancer were determined using highly sensitive SP immunohistochemical method.. Normal immunoreactivity of E-cadherin and beta-catenin were observed in 29 (48.3%) and 18 (30.0%) cases, respectively. Twenty-eight cases (46.7%) showed cyclin D1 overexpression. Thirty percent (9/29) of the cases with normal staining of E-cadherin showed overexpression of cyclin D1, while 61.3%(19/31) of the cases showed overexpression of cyclin D1 with abnormal staining of E-cadherin. Abnormal expression of E-cadherin and overexpression of cyclin D1 showed a significantly positive correlation (rs=0.303,P< 0.05). Forty-two cases showed abnormal staining of beta-catenin. Cyclin D1 overexpression was observed in 57.1% (24/42) of these cases with abnormal staining of beta-catenin, but only observed in 22.2% (4/18) of these cases with normal membranous staining of beta-catenin. There was a significantly positive correlation between the abnormal expression of beta-catenin and overexpression of cyclin D1 (rs=0.321, P< 0.05).. Down-regulation of E-cadherin and beta-catenin accumulation in the cytoplasm/nuclear may promote malignant transformation and progression by triggering cyclin D1 overexpression in breast cancer.

Topics: Adult; Aged; beta Catenin; Breast Neoplasms; Cadherins; Carcinoma, Ductal, Breast; Cyclin D1; Cytoskeletal Proteins; Female; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Middle Aged; Trans-Activators

The aim of this study was to assess the levels of cell cycle regulatory proteins p21waf1 (p21), p53, Cyclin A, Cyclin D1 and Ki-67 to see whether they correlated with recurrence-free survival (RFS). From 1982 to 1996, 50 patients aged less than 51 years underwent lumpectomy followed by radiotherapy for a pure ductal carcinoma in situ (DCIS). For each case, the following immunohistochemical stains were carried out: Ki-67, Cyclin A, Cyclin D1, p53 and p21waf1 (p21). The percentage of positive nuclei was assessed. Multiple combinations of these factors were performed; in particular, we called the sum of Ki-67 and Cyclin A a global proliferation factor (GPF). Correlations with classical clinicopathological data were assessed. After a multivariate analysis, only GPF, Van Nuys Prognostic Index (VNPI) grade and mitotic index were independent predictive factors of recurrence in the whole population. In the population with close surgical margins, when the GPF level was less than the 25th percentile or more than the 75th percentile recurrence was low. In this preliminary study, GPF seems to be of interest to help in the decision process in the post-surgical management of the patient.

Topics: Adult; Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Cycle Proteins; Cyclin A; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p21; Cyclins; Disease-Free Survival; Female; Humans; Immunohistochemistry; Ki-67 Antigen; Middle Aged; Prognosis; Tumor Suppressor Protein p53

B-cell lymphoma gene (BCL-6) upregulation contributes to immortalization of mouse embryo fibroblast and primary B cells via upregulation of cyclin D1. As cyclin D1 overexpression is a common phenomenon in different cancers, BCL-6 protein overexpression may not be restricted to lymphomas. In this study, expression of BCL-6 was investigated by immunohistochemistry on paraffin-embedded specimens from 150 breast cancer patients and 10 specimens of normal breast tissue. The results showed BCL-6 overexpression (> or =10% of cells) in 24/150 (16%) breast cancer patients, whereas in normal breast low expression (<1%) of BCL-6 was observed. In linear regression analysis BCL-6 expression was associated with cyclin D1 (r=0.197, P=0.016). Further, in chi2 analyses, BCL-6-positivity was associated with overexpression of p53 (P=0.016), and hypoxia-inducible factor-1alpha (P<0.001). Involvement of BCL-6 in breast carcinogenesis is further underscored by comparative genomic hybridization analysis that showed gains at the BCL-6 locus (3q27) in 14/86 (16%) breast cancer tissues. The cases with amplification in BCL-6 showed an increased (25%) incidence of BCL-6 protein overexpression. Thus, this study is the first to show that BCL-6 oncogene activation plays a role in cancers other than lymphomas.

Topics: Breast Neoplasms; Carcinoma, Ductal; Carcinoma, Ductal, Breast; Cyclin D1; DNA-Binding Proteins; Female; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; Immunohistochemistry; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-bcl-6; Transcription Factors

All malignant cells appear to have increased needs for glucose in vitro as well as in vivo. The enhanced glucose uptake is mediated through transporters (Gluts), whose action and expression are regulated by oncogenes and growth factors. Cyclin D1 is a nuclear protein that plays an important role in regulating the cell cycle by promoting entry of cells from the G1 to S phase. Increased expression of Glut1 glucose transporter and cyclin D1 have been reported in several neoplasms. In the present study we examined the expression of Glut1 and cyclin D1 in breast carcinomas with negative lymph nodes. We studied 78 infiltrating ductal carcinomas (25 grade 1, 36 grade 2, and 17 grade 3) with negative lymph nodes. Glut1 was expressed in 28% of grade 1, 63.8% of grade 2 and 58.7% of grade 3 carcinomas. Nuclear expression of cyclin D1 was detected in 32% of grade 1, 44.4% of grade 2, and 41.2% of grade 3 carcinomas. From our study it appears that in breast carcinomas with negative lymph nodes Glut1 expression is better correlated to the grade of the neoplasm than cyclin D1.

Topics: Biomarkers, Tumor; Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Female; Glucose Transporter Type 1; Humans; Immunohistochemistry; Lymph Nodes; Lymphatic Metastasis; Monosaccharide Transport Proteins; Predictive Value of Tests

Because of the relatively low incidence of lobular breast carcinoma, there are very few studies on the molecular characteristics of this breast cancer. In an attempt to improve its characterization, we investigated in a large collection of invasive lobular carcinomas (ILCs) the status of markers known to be involved in the better-studied invasive ductal carcinomas (IDC). In the current study we disposed of 80 well-characterized ILC cases. Gene amplification of cyclin D1 (CCND1) and c-erbB2-encoding gene (ERBB2) and expression of their gene products were studied by differential polymerase chain reaction (PCR) and immunohistochemistry, respectively. A comprehensive point mutation study of the phosphatase and tensin homolog tumor suppressor gene (PTEN) was pursued by single strand conformation polymorphism (SSCP)/sequencing analysis. The CCND1 gene was rarely amplified in ILC in spite of showing overexpression of the protein in 41% of tumors. Hence, unlike IDC, increase in gene dosage did not account for the protein excess. PTEN mutations were detected in ILC (truncating mutations) in around 2% of the tumors. Unlike IDC, ILC did not display ERBB2 overexpression and expression of the transcription factor E2F1 correlated inversely with tumor grade. The observed discrepancy in the pattern of the human oncogenes CCND1 and ERBB2, which are involved in the process of carcinogenesis of ductal tumors, appears to suggest a different molecular basis for development and progression of ILC.

Topics: Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Lobular; Cell Cycle Proteins; Cyclin D1; DNA Mutational Analysis; DNA-Binding Proteins; E2F Transcription Factors; E2F1 Transcription Factor; Female; Gene Amplification; Gene Expression Regulation, Neoplastic; Genes, Tumor Suppressor; Humans; Ki-67 Antigen; Neoplasm Invasiveness; Phosphoric Monoester Hydrolases; Point Mutation; Polymorphism, Single-Stranded Conformational; PTEN Phosphohydrolase; Receptor, ErbB-2; Transcription Factors; Tumor Suppressor Proteins

Overexpression of oncogenic proteins may be caused by gene amplifications. Cyclin D1 participates in regulation of the cell cycle. Relations between cyclin D1 expression and amplification of CCND1 gene encoding this protein in invasive duct breast carcinomas (IDC) are not fully elucidated. An increased interest is also focused on relations to the estrogen receptor (ER alpha).. We investigated copy numbers of the CCND1 gene, expression of cyclin D1 and expression of ER alpha in a group of 60 females and 1 male with IDC. The age range varied from 33 to 89 years (median 57 years). The number of CCND1 gene copies and the number of chromosome 11 was evaluated using FISH, the expression of cyclin D1 and ER alpha was investigated by IHC. We detected a strong amplification of CCND1 gene (> 10 copies per tumor cell nuclei) in 9 patients, weak amplification (< or = copies) in 16 patients. Amplification of the CCND1 gene correlated well with the overexpression of cyclin D1. We observed the overexpression of cyclin D1 also in 13 of 36 patients without the gene amplification; therefore, the mechanism of the protein overexpression is different than that caused by the gene amplification in a proportion of patients. Amplification of the CCND1 gene was associated with a high histologic grade of IDC, whereas cases with cyclin D1 overexpression only were not. 24 of 31 patients with overexpression of cyclin D1 coexpressed ER alpha. We did not find correlation between expression/amplification of cyclin D1/CCND1 gene and the size of carcinomas and with metastases to the axillary lymph nodes.. Amplification of the CCND1 gene is associated with overexpression of cyclin D1 in a majority of IDC. Overexpression of cyclin D1 is related to an increased expression of ER alpha. Interaction between cyclin D1 and ER alpha may explain low response to anti-estrogen therapy of some patients.

Topics: Adult; Aged; Aged, 80 and over; Breast Neoplasms; Breast Neoplasms, Male; Carcinoma, Ductal, Breast; Chromosomes, Human, Pair 11; Cyclin D1; Estrogen Receptor alpha; Female; Gene Amplification; Humans; In Situ Hybridization, Fluorescence; Male; Middle Aged; Receptors, Estrogen

Frizzled-related protein (Frp) is a new family of secreted proteins that contain a region homologous to the extracellular cysteine-rich domain (CRD) of the frizzled family proteins. The role of Frp protein is far from clear. To explore the role of Frp and its relationship to the Wnt-signalling pathway in breast cancer, in situ hybridization and immunohistochemical analyses of Frp, Wnt-1, APC, beta-catenin, and its target genes c-myc and cyclin D1 were conducted in 70 specimens of invasive ductal carcinomas of the human breast. Frp mRNA was down-regulated in 62 and elevated in eight tumour specimens, compared with adjacent normal tissues. In the course of tumour progression, however, Frp mRNA steadily increased in both tumour and the adjacent tissues. Interestingly, the number of cases with axillary lymph node metastasis was significantly lower in the group with elevated Frp than in the group with decreased Frp, suggesting that Frp may contribute as a prognostic factor in invasive breast cancer. Wnt-1, a gene implicated in human breast cancer, was markedly elevated in grade 1 tumours, but declined as tumour grade declined. The level of Wnt-1 was linearly correlated with its downstream target beta-catenin (p<0.05), but was inversely correlated with Frp (p<0.05), suggesting a possible negative regulatory role of Frp with regard to Wnt-1. APC was inversely correlated with beta-catenin (p<0.05). Beta-catenin, a key transcriptional activator responsible for the activation of both c-myc and cyclin D1 in colorectal tumours, was detected at high levels in the plasma membranes of cells in normal tissue. In tumour masses, however, beta-catenin lost its tight association with the membrane and diffused into the cytoplasm. Surprisingly, it clearly did not penetrate the nuclei, despite the fact that both c-myc and cyclin D1 were markedly elevated in all tumour tissues. As revealed in this study, Wnt-1/beta-catenin plays very different roles in the oncogenesis of breast and colon cancers. This first systemic analysis of the Frp and the Wnt-signalling pathway in human breast cancer provides a springboard for further work on the role of Frp in the development of breast cancer.

Topics: Adenomatous Polyposis Coli Protein; beta Catenin; Breast; Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Membrane; Cyclin D1; Cytoskeletal Proteins; Female; Follistatin-Related Proteins; Glycoproteins; Humans; Immunohistochemistry; In Situ Hybridization; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-myc; Signal Transduction; Trans-Activators; Wnt Proteins; Wnt1 Protein; Zebrafish Proteins

CyclinD1 plays a critical role in regulating cell cycle progression. CyclinD1 mRNA and protein are overexpressed in approximately 50% of primary breast cancer cases. However, its clinical significance as a predictive factor remains unclear. One hundred and seventy-three female patients diagnosed with invasive ductal carcinoma who had undergone a mastectomy (161 patients) or breast-conserving surgery (12 patients) were followed up for 6-119 months (median 86 months) postoperatively. Immunoreactivity for monoclonal anti-cyclinD1 antibody (clone DCS-6) with paraffin-embedded carcinoma tissues was investigated using a labeled streptavidin-biotin method. Overexpression of cyclinD1 was found in 42% (73 of 173), and strongly correlated with estrogen receptor (ER) expression (p < 0.000001). Univariate analysis revealed no association between overexpression of cyclinD1 and overall survival or relapse-free survival in all patient groups. However, in the ER-negative subgroup (n = 75), overexpression of cyclinD1 was significantly correlated with shorter overall survival (p = 0.018) and relapse-free survival (p = 0.014) as well as the lymph node status and tumor size. In contrast, there were no significant associations between overexpression of cyclinD1 and clinical outcome in the ER-positive subgroup. According to Cox's multivariate analysis in the ER-negative subgroup, overexpression of cyclinD1 had the most significant effect on overall survival (p = 0.02) and relapse-free survival (p = 0.0058), followed by nodal status and histologic grade. These findings suggest that overexpression of cyclinD1 is an independent prognostic indicator in ER-negative breast cancer patients.

Topics: Adult; Aged; Aged, 80 and over; Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Female; Humans; Immunoenzyme Techniques; Lymph Nodes; Mastectomy; Middle Aged; Neoplasm Staging; Prognosis; Receptors, Estrogen; Receptors, Progesterone; Survival Rate; Treatment Outcome

To account for the accumulation of genomic alterations required for tumor progression, it has been suggested that the genomes of cancer cells are unstable and that this instability results from defective mutators (the "mutator phenotype" theory). To examine the hypothesis that abnormal cell-cycle regulators act as the mutators contributing to genomic instability, the present study, based on primary tumor tissues from 71 patients with breast cancer, was performed to determine whether there was an association between aberrant expression of cell-cycle regulators (cyclin A, cyclin D1, cyclin E, RB1, p21, and p27) and chromosomal instability. Comparative genomic hybridization was used to measure chromosomal changes, reflecting genomic instability in individual tumors, whereas immunohistochemistry was used to detect aberrant expression of cell-cycle regulators. Overexpression of cyclin D1 was found to be significantly correlated with increased chromosomal instability (defined as harboring more than 7 chromosomal changes), with 63% of tumors overexpressing and 27% of tumors not overexpressing, with cyclin D1 showing chromosomal instability (P < 0.05). Interestingly, this relationship was independent of cell outgrowth (as detected by the proliferation marker Ki-67) and was particularly significant in tumors not expressing p27 or in tumors with detectable RB1. These results suggest that cyclin D1 plays an alternative role in the regulation of genomic stability.

Topics: Adult; Aged; Aged, 80 and over; Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Cycle Proteins; Chromosome Aberrations; Chromosome Deletion; Cyclin D1; Female; Gene Amplification; Genome, Human; Humans; Middle Aged; Nucleic Acid Hybridization

To understand with greater clearness the effect of overexpression of cyclins gene and the potential implications of it for tumorgenesis in breast.. We assayed the expression of cyclins D1, E and A in different types of human breast diseases by immunohistochemical staining.. Significant difference was seen among the malignant tumor, benign tumor and dysplasis(P < 0.001). The expressive intensity of above three kind of cyclins in malignant tumor was the highest, that in benign tumor was higher, and that in dysplasis was low.. These data suggest that the expressive intensity of cyclins gene may serve as an indicator for malignant intensity of tumors. The expressive ratio difference of cyclins E and A between infiltrating ductal carcinoma and infiltrating lobular carcinoma may imply that there are different mechanisms involving the occurrence of different histological types of breast tumor.

Topics: Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin A; Cyclin D1; Cyclin E; Female; Humans

In order to understand the intricate relationship of cell proliferation and apoptosis in tumour development, proliferation markers (Ki-67 and c-myc), apoptosis, cell-cycle inducers cyclin D1 and D3, and cell-cycle inhibitors p16(INK4), p21(CIP1), and p27(KIP1) were evaluated in ductal breast carcinoma. The heterogeneous nature of breast tumours provides a system by which the changes in cell-cycle genes can be explored under a wide range of proliferation and apoptotic indices. To address the above issues, immunohistochemical studies were conducted in 40 pairs of tumours and adjacent normal ductal tissues. The TUNEL method was used to identify apoptotic cells. Except for p27/KIP1, the proliferation (Ki-67, c-myc) and the apoptotic indexes together with levels of p16/INK4a, p21/CIP1, cyclin D1, and cyclin D3, were clearly elevated among tumour tissues, while absent in the adjacent normal tissues. Spearman correlation analysis indicated strong associations among apoptotic index, Ki-67, c-myc, and tumour grade. In addition, p21/CIP1 and cyclin D3 were positively correlated, while p16/INK4a, p27/KIP1, and cyclin D1 were negatively correlated with tumour grade. There was clear decoupling between p21 and p27, as well as decoupling between cyclin D1 and cyclin D3, in terms of their relationship to cell proliferation and apoptosis, indicating differential roles in tumour progression.

Topics: Apoptosis; Biomarkers, Tumor; Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Cycle Proteins; Cell Division; Cyclin D1; Cyclin D3; Cyclin-Dependent Kinase Inhibitor p16; Cyclin-Dependent Kinase Inhibitor p21; Cyclin-Dependent Kinase Inhibitor p27; Cyclins; Disease Progression; Female; Humans; In Situ Nick-End Labeling; Microtubule-Associated Proteins; Neoplasm Proteins; Tumor Suppressor Proteins

There is increasing evidence that there are different progression routes leading to invasive breast cancer, depending on histology and differentiation grade. The aim of this study was to determine alterations in the expression of proteins involved in proliferation and apoptosis in non-invasive and invasive ductal breast lesions. Immunohistochemistry was performed on 106 usual ductal hyperplasias (UDH), 61 DCIS lesions and 53 invasive ductal breast carcinomas. Increased proliferation (Ki67), overexpression of cyclin D1, HER-2/neu, p21 and p53, and decreased expression of bcl-2 and p27 could already be found in UDH. Significant differences between UDH and DCIS lesions were found for only one protein when UDH was compared with well-differentiated DCIS (p27), for three proteins when compared with intermediately differentiated DCIS (p21, cyclin D1, Ki-67), and for all proteins when compared with poorly-differentiated DCIS. Comparing DCIS with invasive lesions of same differentiation grade, proliferation was elevated in the invasive lesions. Altered expression of the other proteins was in general only slightly increased in the invasive lesion compared with DCIS. The number of proteins with altered expression per lesion was highest in poorly-differentiated lesions and was comparable between DCIS and invasive cancer of the same differentiation grade. In conclusion, the biggest changes in expression of these proliferation and apoptosis related proteins appear to occur during the transition from hyperplasia to DCIS; they probably play a minor role in the transition from DCIS to invasive breast lesion of same differentiation grade. Well-differentiated in situ and invasive breast lesions share many of the aberrations in expression of these proteins, as do poorly-differentiated in situ and invasive lesions. However, there are many differences between the well and poorly-differentiated lesions. This further supports the existence of different progression routes leading to breast cancer.

Topics: Apoptosis; Breast Neoplasms; Carcinoma in Situ; Carcinoma, Ductal, Breast; Cell Cycle Proteins; Cyclin D1; Female; Humans; Immunohistochemistry; Ki-67 Antigen; Proto-Oncogene Proteins c-bcl-2; Receptor, ErbB-2; Tumor Suppressor Protein p53

Human solid tumors undergo clonal evolution as they progress, but evidence for specific sequences of genetic changes that occur in individual tumors and are recapitulated in other tumors is difficult to obtain.. Patterns of amplification of Her-2/neu, c-myc, and cyclin D1 were determined by fluorescence in situ hybridization (FISH) in relation to the presence of p53 dysfunction and ploidy in 60 primary human breast cancers.. We show that there are clusters of genophenotypic abnormalities that distinguish lobular breast cancers from nonlobular tumors; that cyclin D1 amplification occurs prior to the divergence of lobular breast cancers from nonlobular cancers; that p53 dysfunction, Her-2/neu amplification, and c-myc amplification are characteristic features of nonlobular breast cancers, but not of lobular breast cancers; and that the frequencies of amplification of all three oncogenes examined increase progressively with increasing aneuploidy, but that each gene exhibits a different profile of increasing amplification in relation to tumor progression. Early amplification of c-myc appears to be an especially prominent feature of hypertetraploid/hypertetrasomic tumors.. The data suggest that in tumors containing multiple abnormalities, these abnormalities often accumulate in the same cells within each tumor. Furthermore, the same patterns of accumulation of multiple genophenotypic abnormalities are recapitulated in different tumors.

Topics: Alleles; Aneuploidy; Breast Neoplasms; Carcinoma, Ductal, Breast; Chromosomes, Human, Pair 17; Cyclin D1; Disease Progression; Genes, p53; Genotype; Humans; In Situ Hybridization, Fluorescence; Loss of Heterozygosity; Phenotype; Ploidies; Proto-Oncogene Proteins c-myc; Receptor, ErbB-2

Infiltrating ductal mammary carcinomas are histologically graded according to their extent of differentiation. Well-differentiated, grade I, tumours have low proliferative activity, usually form tubules and exhibit little nuclear pleomorphism. Despite an apparently reassuring morphology, 10-15% of grade I ductal carcinomas metastasize, albeit after a prolonged period. Recent evidence supports the view that evolution to higher grade malignancies occurs rarely and that grade I tumours are biologically distinct from grade III tumours. We have examined a series of 148 grade I ductal carcinomas in order to ascertain whether information about the level of expression of cyclin D1, p27, p53, oestrogen receptor status (ER) or proliferative activity could be used to identify those patients with a poor outcome. The majority of tumours expressed high levels of cyclin D1, p27 and ER, low levels of p53 and had low Ki-67 expression and mitotic counts. Cyclin D1, p27 and ER expression were all significantly correlated with each other but not with p53 (cyclin D1 correlation with ER, p = 0.01; cyclin D1 correlation with p27 and ER correlation with p27 both p < 0.0001). Cyclin D1 and ER were also both correlated with Ki-67 (p = 0.01 and p < 0.0001) but not with mitotic count. Our results suggest that cyclin D1, ER and p27 are all markers of well-differentiated tumours and that their detection is related to proliferative activity in a manner reflecting their functional role within the normal cell cycle. However, none of the proteins or markers of proliferative activity were sensitive enough to predict which patients were likely to have a poor outcome.

Topics: Adult; Aged; Aged, 80 and over; Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Cycle Proteins; Cell Division; Cyclin D1; Female; Humans; Immunohistochemistry; Ki-67 Antigen; Microfilament Proteins; Middle Aged; Mitotic Index; Muscle Proteins; Phenotype; Receptors, Estrogen; Tumor Suppressor Protein p53

The cell cycle regulatory gene, Cyclin D1, plays a critical role in the growth and progression of several types of human cancer, including breast cancer. Immunohistochemical study of Cyclin D1 expression has been extensively reported in invasive ductal carcinoma (IDC). In contrast, there have been few reports concerning Cyclin D1 expression in ductal carcinoma in situ (DCIS) and their positive rates are variable. The differences in the reported frequency may be largely due to the differences in antibodies used, immunohistochemical methods and the positive cut-off point. However, we speculated that the strictness of diagnosis of DCIS might be somewhat responsible for these differences in frequency. Therefore, we selected cases of DCIS by carefully eliminating cases of predominantly intraductal carcinoma (PIC). Moreover, to clarify whether Cyclin D1 expression is involved in multistep carcinogenesis or the progression of human breast cancer, we immunohistochemically investigated Cyclin D1 expression in 57 DCIS, 10 atypical ductal hyperplasia (ADH), 70 usual ductal hyperplasia (UDH), 44 PIC and 92 IDC. Cyclin D1 expression was detected in 41 DCIS cases (72%), 22 PIC cases (50%) and 40 IDC cases (43%). No expression of Cyclin D1 was observed in either ADH or UDH. There were no significant correlations between Cyclin D1 expression and histological grade or estrogen receptor expression in DCIS. These results suggest that Cyclin D1 expression may play an important role in the early stages of carcinogenesis, and that immunohistochemical detection of Cyclin D1 expression may be helpful in differentiating low-grade DCIS from ADH.

Topics: Breast Neoplasms; Carcinoma in Situ; Carcinoma, Ductal, Breast; Cyclin D1; Female; Fluorescent Antibody Technique, Indirect; Humans; Hyperplasia; Immunoenzyme Techniques; Lymph Nodes; Lymphatic Metastasis; Neoplasm Staging; Receptor, ErbB-2; Receptors, Estrogen; Tumor Suppressor Protein p53

Breast cancer is probably the result of a series of genetic events, each with its own histopathologic correlate in the hyperplasia to carcinoma sequence. The expression of breast cancer markers in hyperplasia and tumors are well known, but few studies have investigated their sequential expression among hyperplastic and cancerous lesions within the same breast. Using breast tissue obtained from a single procedure, we correlated the immunohistochemical expression of several breast cancer markers with the histopathologic stage of proliferative breast disease. We selected 14 cases in which various degrees of hyperplasia coexisted with carcinoma. Serial sections were reacted with antibodies to DF3, c-erbB-2, p53 (DO7 and CM1), B72.3, and cyclin D1. We found that within an individual breast, the number of breast cancer markers expressed increased with progression from hyperplasia to atypical hyperplasia to carcinoma. Cytoplasmic DF3 was first expressed at the level of simple hyperplasia, followed by c-erbB-2 in atypical hyperplasia. Overexpression of p53 was confined to carcinomas, and thus appeared to be a late event. B72.3 was expressed in three carcinomas and in one atypical hyperplasia, although the associated carcinoma was negative. Carcinomas that expressed cytoplasmic DF3 and c-erbB-2 were associated with atypical hyperplasias that also expressed cytoplasmic DF3 and c-erbB-2, with one and two exceptions, respectively. No specific cyclin D1 staining pattern was observed.

Topics: Antibodies; Antigens, Neoplasm; Biomarkers, Tumor; Breast; Breast Neoplasms; Carcinoma in Situ; Carcinoma, Ductal, Breast; Carcinoma, Lobular; Cyclin D1; Female; Glycoproteins; Humans; Hyperplasia; Immunohistochemistry; Receptor, ErbB-2; Tumor Suppressor Protein p53

Infiltrating lobular carcinoma (ILC) and infiltrating ductal carcinoma (IDC) are similar in many respects and their histologic features occasionally overlap. Despite the many similarities, some clinical follow-up data and the patterns of metastasis suggest that ILC and IDC are biologically distinct. Unfortunately, most breast cancer research has focused almost exclusively on the ductal subtype or has not stressed the biologic or molecular genetic distinctions between breast carcinoma subtypes. Several reports have suggested the possibility that ILCs and IDCs differ with respect to expression of antigens involved in proliferation and cell cycle regulation. Therefore, we undertook an immunohistochemical evaluation of cell cycle related antigens in ILCs, including histologic variants thought to represent aggressive neoplasms, and IDCs matched for histologic grade (Modified Bloom-Richardson Grade I). We believe that different antigen expression profiles could elucidate the biological distinctiveness of breast carcinoma subtypes and possibly provide diagnostically relevant information. We studied the expression of the following antigens in 28 archived, formalin-fixed ILCs and 34 well-differentiated IDCs: estrogen receptor (ER), progesterone receptor (PR), Her 2-neu, mib-1, cyclin D1, p27, p53, mdm-2 and bcl-2. 94% of ILCs and 100% of IDCs expressed ER; 75% of ILCs and 76% of IDCs expressed PR; 4% of ILCs and 13% of IDCs expressed c cerb B-2; ILCs and IDCs both expressed mib-1 in approximately 10% of lesional cells; 82% of ILCs and 54% of IDCs expressed cyclin D1; 90% of ILCs and 83% IDCs expressed p27 strongly; 4% of ILCs and 4% of IDCs expressed p53, 25% of ILCs and 33% of IDCs expressed mdm-2; 96% of ILCs and 100% of IDCs expressed bcl-2. None of the apparent differences were statistically significant. The ILC variants demonstrated immunophenotypes that were essentially similar to ILCs of the usual type. We conclude that ILCs and well-differentiated IDCs show similar proliferation and cell cycle control antigen profiles. Despite their unusual histologic features, most ILC variants appear to maintain a characteristic ILC immunophenotype.

Topics: Antigens, Neoplasm; Antigens, Nuclear; Biomarkers; Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Lobular; Cell Cycle; Cell Cycle Proteins; Cohort Studies; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p27; Female; Humans; Immunophenotyping; Ki-67 Antigen; Microtubule-Associated Proteins; Neoplasm Invasiveness; Neoplasm Metastasis; Neoplasm Proteins; Nuclear Proteins; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-bcl-2; Proto-Oncogene Proteins c-mdm2; Receptor, ErbB-2; Receptors, Estrogen; Receptors, Progesterone; Tumor Suppressor Protein p53; Tumor Suppressor Proteins

In a previous study, we demonstrated that the G1 cell cycle checkpoint in carcinomas of the breast is frequently abrogated by loss of p16, the product of the CDKN2/INK4A gene, and, to a lesser extent, by loss of pRB, the product of the retinoblastoma gene. The purpose of the present study was to determine whether other mechanisms of cell cycle deregulation exist in breast cancers which have retained RB and p16 function. Paraffin sections of 81 invasive breast carcinomas (49 ductal, 26 lobular, 6 mixed) were reacted with monoclonal antibodies against cyclin D1 and p53, using optimized immunohistochemical staining protocols. The staining results were correlated with the expression of p16 and pRB, and with a variety of pathological parameters and DNA ploidy. Twenty-five tumors (31%) accumulated (presumably mutant) p53 and 28 (35%) overexpressed cyclin D1; 7 carcinomas (not including any pure lobular cancers) abnormally expressed both proteins. p53 accumulation correlated with nuclear, mitotic, and overall grade, but not with tumor size, lymph node involvement, or DNA ploidy. Overexpression of cyclin D1 was not associated with any of the patho-biological variables. There was an inverse correlation between loss of p16 and high levels of p53, but not cyclin D1. The G1 cell cycle checkpoint, which is controlled by RB, cyclin D1, and p16, was abrogated in 65% of carcinomas, and only p53 was abnormal in an additional 17%. The number of abnormally expressed genes correlated with mitotic activity and overall tumor grade, but not with tumor histology, size, or nodal status, suggesting that cell cycle deregulation is an early event in breast tumorigenesis. Only 18% of the carcinomas showed a normal level of expression of the four genes tested, and p16 appeared to be the most common target of cell cycle deregulation. These data point to the importance of cell cycle regulatory protein abnormalities in human breast cancer.

Topics: Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Lobular; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; DNA, Neoplasm; Female; Humans; Immunoenzyme Techniques; Mutation; Ploidies; Tumor Suppressor Protein p53

Investigation of early breast carcinogenesis is limited by the difficulty in obtaining cell cultures or adequate fresh frozen material and by the fact that available data from in situ techniques are interpreted in terms of various classification systems. Our studies in a series of pure ductal carcinomas in situ (DCIS) were conducted in accordance with the recommendations of the international Consensus Conference (Hum. Pathol., 28, 122-125, 1997) relative to processing, determination of lesion extent, and histological stratification primarily on nuclear grade (NG). A multifactorial study performed in 15 low- and 16 high-NG DCIS (68% detected by mammography) included the following: (1) morphological analysis of NG, necrosis, and architectural pattern; (2) detection of numerical genomic abnormalities at ERBB2, MYC, CCND1, Xq1.2 and 20q13 loci by fluorescence in situ hybridization on interphase nuclei; and (3) immunohistochemical determination of cell proliferation, p53 accumulation, hormonal receptors and bcl-2 expression on serial sections of formalin-fixed, paraffin-embedded specimens. High NG, comedo/solid pattern and necrosis were significantly associated with amplification at one or more loci, the number of amplified loci, amplification at the ERBB2 locus, absence of bcl-2 and hormonal receptor expression and high cell proliferation (p < 0.05). High NG and comedo/solid pattern were significantly associated with MYC amplification and p53 accumulation, and necrosis with CCND1 amplification (the only gene amplification detected in low NG DCIS). These data provide additional information on the early steps of breast carcinogenesis, in accordance with currently recognized criteria of histological classification.

Topics: Adult; Aged; Breast Neoplasms; Carcinoma in Situ; Carcinoma, Ductal, Breast; Cell Division; Cyclin D1; Female; Gene Amplification; Genes, erbB-2; Genes, myc; Humans; Immunohistochemistry; In Situ Hybridization, Fluorescence; Middle Aged; Proto-Oncogenes

Gene amplification is one essential mechanism leading to oncogene activation which is supposed to play a major role in the pathogenesis of invasive breast cancer. However, using standard methodologies the detection of gene amplifications has been limited especially in small-sized lesions, like pre-invasive precursor lesions. The combination of two novel technologies, laser-based microdissection and quantitative real-time PCR, facilitates the detection of low-level amplifications in morphologically defined lesions. As a model system we investigated in situ breast cancer (ductal carcinoma in situ, DCIS) classified according to the morphology-based Van Nuys grading system for amplification of growth-regulatory genes. In this study 83 formalin-fixed, paraffin-embedded archival DCIS specimens were examined after laser-based microdissection by quantitative real-time PCR using the TaqMan detection system for amplification of the c-erbB2, topoisomerase IIalpha, c-myc and cyclinD1 gene. In a subset of 17 DCIS with adjacent infiltrating tumour components we compared intraductal and invasive tumour components in parallel for differences in amplification status. The combination of these new techniques represents an excellent tool to gain new insights into carcinogenesis by analyzing genetic alterations in morphologically identified heterogeneous lesions in breast cancer progression within the very same specimen or even tissue slide.

Topics: Antigens, Neoplasm; Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Intraductal, Noninfiltrating; Cell Separation; Cyclin D1; Dissection; DNA Topoisomerases, Type II; DNA-Binding Proteins; Female; Gene Amplification; Genotype; Humans; Isoenzymes; Lasers; Phenotype; Polymerase Chain Reaction

Cyclin-D1 (CD1) expression was analyzed in human mammary carcinomas by immunohistochemical (IHC) and flow-cytometry (FCM) methods: 52.5% and 50% of cases were strong expressors of CD1 by IHC and FCM analysis respectively. The percentage of CD1-positive cells was especially high in node-negative (N-) estrogen-receptor-positive (ER+) tumors, probably as a consequence of CD1 induction by estrogens in steroid-responsive tissues. However, CD1 expression was not related to ER positivity in node-positive tumors (N+). An interesting relationship between CD1 expression and H3-thymidine labelling index (H3Td-LI) was also found: CD1 and H3Td-LI were unrelated in N- tumors, while high CD1 expression was observed in N+ tumors with high DNA synthesis, as assessed by H3Td-LI. The combined measurement of DNA and CD1 showed that 27 specimens were aneuploid, 19 of them (19/27; 70%) strongly expressing CD1. Further studies are needed to clarify the role of CD1 in DNA abnormality of breast tumors. However, we cannot exclude that the CD1 may be differently de-regulated in the last phase of tumor progression, and that CD1 over-expression may contribute to the aneuploidy of mammary carcinomas.

Topics: Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Division; Cyclin D1; Female; Flow Cytometry; Humans; Immunohistochemistry; Lymphatic Metastasis; Middle Aged; Neoplasm Proteins; Ploidies; Receptors, Estrogen

The restriction of energy intake has a profound inhibitory effect on carcinogenesis, yet the mechanism or mechanisms that account for this effect are unknown. In this experiment, the hypothesis tested was that energy restriction upregulates the expression of p27/kip1, a gene product associated with cell-cycle growth arrest, while downregulating cyclin D1, a protein that combines with cyclin-dependent kinases to promote phosphorylation of retinoblastoma protein and the progression of cells through the cell cycle. We studied levels of these proteins in uninvolved mammary epithelial cells and in mammary intraductal proliferations, ductal carcinomas in situ, and adenocarcinomas induced in response to administration of 1-methyl-1-nitrosourea in animals fed either ad libitum or 90%, 80%, or 60% of ad libitum intake. Protein levels were evaluated immunohistochemically by using computer-assisted image analysis to quantify differences in protein expression among treatment groups. The expression of p27 increased and the expression of cyclin D1 decreased dose-dependently in response to energy restriction. The effect was greater on p27 than on cyclin D1. The hypothesis proposed is that energy restriction inhibits carcinogenesis by arresting cell-cycle progression by regulating p27/kip1.

Topics: Adenocarcinoma; Animals; Carcinoma in Situ; Carcinoma, Ductal, Breast; Carcinoma, Intraductal, Noninfiltrating; Cell Cycle; Cell Cycle Proteins; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p27; Energy Intake; Energy Metabolism; Female; Food Deprivation; Gene Expression Regulation, Neoplastic; Mammary Glands, Animal; Mammary Neoplasms, Experimental; Methylnitrosourea; Microtubule-Associated Proteins; Neoplasm Proteins; Precancerous Conditions; Rats; Rats, Sprague-Dawley; Tumor Suppressor Proteins

Cyclin D1 (CCND1) amplification is found in 10-15 per cent of invasive breast carcinomas, but it is not well established whether this gene alteration also occurs in the precursor of invasive breast carcinoma, ductal carcinoma in situ (DCIS). By Southern blot analysis, cyclin D1 gene amplification was detected in 10 per cent (3/32) of DCIS cases. In addition, 15 cases of DCIS were analysed using bright field in situ hybridization (BRISH), of which 11 had already been analysed by Southern blotting. One additional case with gene amplification was found by BRISH. The use of BRISH for the detection of gene amplification is shown to be a novel and reliable in situ method on paraffin-embedded tissue sections. By immunohistochemistry, 147 cases of DCIS were analysed for the expression of cyclin D1. Cyclin D1 overexpression was found in 9 per cent of well-differentiated, 29 per cent of intermediately differentiated, and 19 per cent of poorly differentiated DCIS. No statistically significant association was found between cyclin D1 overexpression and the differentiation grade of DCIS, although 90 per cent of the cases that show overexpression are classified as intermediately and poorly differentiated. An association was found between cyclin D1 overexpression and oestrogen receptor positivity. Cyclin D1 overexpression was found in all four cases with cyclin D1 gene amplification, but was also found in 30 per cent (8/27) of cases without detectable gene amplification. It is concluded that cyclin D1 gene amplification is an early event in the development of breast carcinoma and occurs in poorly differentiated DCIS. Cyclin D1 protein overexpression is also present in tumours without cyclin D1 gene amplification and is seen predominantly in DCIS of intermediately and poorly differentiated histological type and oestrogen receptor positivity.

Topics: Blotting, Southern; Breast Neoplasms; Carcinoma in Situ; Carcinoma, Ductal, Breast; Cyclin D1; Female; Gene Expression; Genes, bcl-1; Humans; Immunoenzyme Techniques; In Situ Hybridization; Neoplasm Proteins; Receptors, Estrogen

Resistance to mammary tumorigenesis in Copenhagen rats is associated with loss of early preneoplastic lesions known as intraductal proliferations. The cause of this disappearance, however, is unknown.. There were no differences in the numbers of lesions in mammary whole-mounts prepared from Copenhagen or Wistar-Furth rats at 20 or 30 days after N-methyl-N-nitrosourea treatment, but at 37 days there were significantly fewer lesions in Copenhagen glands. Furthermore, lesions in Copenhagen glands were exclusively intraductal proliferations, whereas in Wistar-Furth glands more advanced lesions were also present. Immunohistochemical staining showed frequent cyclin D1 overexpression in Wistar-Furth lesions at 37 days, but not in Copenhagen lesions. There were, however, no differences in p16INK4a protein expression, bromodeoxyuridine labeling and apoptotic indices, or mast cell infiltration between Copenhagen and Wistar-Furth lesions at any time.. Overexpression of cyclin D1 in preneoplastic lesions may be important in the development of mammary tumors in susceptible rats, although this overexpression does not appear to cause significant changes in cell kinetics. Furthermore, the low levels of cyclin D1 expression in Copenhagen intraductal proliferations may play a role in the resistance of these rats to mammary tumorigenesis.

Topics: Animals; Bromodeoxyuridine; Carcinoma in Situ; Carcinoma, Ductal, Breast; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Female; Immunohistochemistry; Mammary Neoplasms, Experimental; Methylnitrosourea; Precancerous Conditions; Rats; Rats, Inbred WF

The cell cycle-associated retinoblastoma protein (pRb) and p16 protein were demonstrated using immuno-histochemistry on paraffin sections from 192 cases of invasive breast carcinoma. Abnormal expression of pRb was defined as negative staining and was seen in 17% of tumours. Such abnormal expression was significantly more frequent in tumours with negative oestrogen receptor (ER) status. There was also a trend for tumours which were negative for pRb to be grade III ductal carcinomas. There was no association between p16 staining and any histopathological parameter, though, surprisingly, log-rank analysis showed that strong staining was associated with a poor outcome. There was a significant inverse relationship between pRb and p16 expression and a significant positive association between pRb and cyclin D1. In a Cox multivariate analysis, which included cyclin D1, neither pRb nor p16 was an independent predictor of patient outcome.

Topics: Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Humans; Immunohistochemistry; Multivariate Analysis; Prognosis; Receptors, Estrogen; Retinoblastoma; Survival Analysis

p21WAF1/Cip1 is an inhibitor of cdk/cyclin complexes, and thus regulates the cell cycle. p21 is also related to cell differentiation and is regulated by wild-type p53, although p53-independent regulatory pathways have been proposed. In order to analyse p21 expression as well as its relationship with p53 in human breast cancer, an immunohistochemical analysis was undertaken of 77 breast carcinomas, 16 of them with an in situ component; 30 adjacent normal tissue samples; and five non-neoplastic specimens. Forty-four infiltrating carcinomas (57 per cent) were p21-positive. Expression of p21 was also observed in pre-invasive lesions, whereas normal ducts were negative or focally and weakly positive. p21 expression was associated with high histological grade (II + III) (P = 0.017) and poor tubule formation (P = 0.002), and was significantly less frequent in lobular carcinomas (P = 0.0001). p21 positivity also correlated with increased proliferation, but this seemed to be dependent on the histological grade. Twenty carcinomas (26 per cent) showed p53 overexpression, but this was not associated with p21 negativity, suggesting the existence of p53-independent mechanisms for p21 regulation in vivo. Cyclin D1CCND1 expression was analysed in the same series and an association between p21 and cyclin D1 expression was found, since 23 of 26 cyclin D1-positive carcinomas were p21-positive (P < 0.001 ...). In conclusion, p21 is frequently overexpressed in breast carcinomas and this occurs in the early stages of neoplastic progression. This overexpression seems to be independent of p53 status and might be involved in cyclin D1 modulation.

Topics: Blotting, Western; Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Lobular; Cell Differentiation; Cell Division; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p21; Cyclins; Disease Progression; Female; Humans; Immunoenzyme Techniques; Neoplasm Proteins; Tumor Suppressor Protein p53

Progression through G1 phase of the eukaryotic cell cycle is tightly controlled by cyclin-dependent kinases (CDK). These proteins form part of a regulatory pathway including the cyclin-dependent kinase inhibitor (CKI) p16, D-type cyclins and the product of the retinoblastoma gene pRb. Aberration of any one of these components may lead to uncontrolled proliferation contributing to neoplasia. Three of these proteins, cyclin D1, pRb and p16, were analysed by immunohistochemistry on archival paraffin sections to determine whether expression patterns were different in preinvasive ductal carcinoma in situ (DCIS) and invasive breast tumours relative to normal. Genetic analysis of the gene encoding cyclin D1 (CCND1) was also carried out, using an intragenic restriction fragment-length polymorphism (RFLP) to assess possible allelic imbalance. A majority of the tumours studied (approximately 90%) showed abnormalities in expression of at least one of these proteins. Overexpression of cyclin D1 was found in approximately 49% cases, reduced expression of p16 in approximately 46% and reduced expression of pRb in approximately 37%. Allelic imbalance of cyclin D1 was found in approximately 57% cases.

Topics: Alleles; Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Cyclin-Dependent Kinases; Female; Humans; Immunohistochemistry; Neoplasm Proteins; Retinoblastoma Protein

Experimental studies suggest that cyclin D1 is a potential oncogene but in clinical studies of invasive breast cancer, overexpression of cyclin D1 is found to be associated with oestrogen receptor (ER) expression and low histological grade, both markers of good prognosis. Immunohistochemistry has been used to examine the relationship between cyclin D1 expression and differentiation in 36 cases of ductal carcinoma in situ (DCIS) and the interrelationship between expression of cyclin D1, its associated protein product of the retinoblastoma gene (pRb), and ER, in this group of cases. The expression of these markers has also been examined in nine cases of atypical ductal hyperplasia (ADH) and these results have been compared with the levels of expression seen in DCIS. Cyclin D1 overexpression was found in 23/36 (64 per cent) cases of DCIS and, in contrast to invasive carcinoma, there was no relationship with either differentiation or ER expression. The level of pRb expression was significantly associated with cyclin D1 expression (rS = 0.49, P = 0.001) and only two cases (6 per cent) were pRb-negative. There was no association between pRb and differentiation of DCIS or ER status. In contrast to DCIS, only one case of ADH showed overexpression of cyclin D1 (Mann-Whitney U-test, P = 0.02). All cases of ADH were ER-positive and showed moderate pRb staining, similar to that seen in well-differentiated DCIS. These results provide further evidence that overexpression of cyclin D1 plays a role early in carcinogenesis.

Topics: Adult; Aged; Aged, 80 and over; Breast; Breast Neoplasms; Carcinoma in Situ; Carcinoma, Ductal, Breast; Cyclin D1; Female; Humans; Hyperplasia; Immunoenzyme Techniques; Middle Aged; Neoplasm Proteins; Precancerous Conditions; Receptors, Estrogen; Retinoblastoma Protein

Cyclin D1 has been reported to be overexpressed in many tumours, including breast carcinomas. Cyclin D1 was first identified as a protooncogene (BCL1/PRAD1), and its overexpression was related to tumour proliferation. The product has also recently been identified as important in mediating cell cycle growth arrest via the p53 pathway in murin fibroblast cell lines. Ninety breast carcinomas previously analysed for p53 status were analysed for amplification of cyclin D1, D2 and D3 genes by Southern blot analysis and for protein expression by immunhistochemistry. In 10 samples gene amplification was detected at the cyclin D1 locus. No gene amplification was detected at the cyclin D2 and D3 loci. Immunoreactivity for cyclin D1 was detected in 38 (42.2%) tumour tissue samples. Fifty samples were immunostained for cyclin D2 and D3. Only 2 samples (4%) showed immunoreactivty for cyclin D2, and 9 samples (18%) for cyclin D3. Cyclin D1 protein overexpression was significantly more often found in tumours with wild type p53 and in tumours with higher grades of differentiation expressing ER. No association was seen between gene amplification of the cyclin D1 gene and p53 status. We conclude there is a relationship between wild type p53 and cyclin D1 protein overexpression in clinical material, indicating that cyclin D1 may be another downstream effector of p53.

Topics: Adult; Aged; Aged, 80 and over; Blotting, Southern; Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Lobular; Cyclin D1; Cyclin D2; Cyclin D3; Cyclins; DNA, Neoplasm; Female; Gene Amplification; Humans; Immunoenzyme Techniques; Middle Aged; Retinoblastoma Protein; Tumor Suppressor Protein p53

Cyclin D1 plays a critical role in regulating cell-cycle progression. Gene amplification and protein overexpression of cyclin D1 have been detected in breast cancer but little is known concerning whether these changes occur in normal breast tissue and in breast lesions associated with increased risk of development of invasive breast cancer. We looked for cyclin D1 gene amplification and protein overexpression in 30 cases of benign breast disease (16 epithelial hyperplasias without atypia and 14 atypical ductal hyperplasias) and 18 ductal carcinomas in situ by use of differential PCR and immunohistochemical staining. We compared the resulting frequencies to those in 15 cases of normal breast tissue and 17 invasive ductal carcinomas. We found cyclin D1 gene amplification in 15% of those with normal breast tissue, 19% of those with epithelial hyperplasia without atypia, 27% of those with atypical ductal hyperplasia, 35% of those with ductal carcinoma in situ, and 25% of those with invasive ductal carcinoma; corresponding figures for protein overexpression were 13, 13, 57, 50, and 64%. These results suggest that cyclin D1 amplification and protein overexpression can occur before histologic alterations are seen but that the frequencies of these changes are higher in histologic lesions with cellular atypia (atypical hyperplasia and ductal carcinoma in situ), reaching frequencies similar to those observed in invasive carcinoma.

Topics: Breast; Breast Diseases; Breast Neoplasms; Carcinoma in Situ; Carcinoma, Ductal, Breast; Cyclin D1; DNA, Neoplasm; Gene Amplification; Gene Expression; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Tumor Cells, Cultured

Expression of proliferation- and apoptosis-related proteins was studied by immunohistochemistry in 130 usual ductal hyperplasias of the breast, of which 39 cases (30%) had adjacent invasive cancer. Overexpression of cyclin D1 and Ki-67 was found in 6% and 29% of the cases, respectively. Only two mild ductal hyperplasias were Her-2/neu positive. Overexpression of p21 and reduced expression of p27, both cdk-inhibitors, was seen in 16% and 27% of the lesions, respectively. Reduced expression of bcl-2 was found in 16% of the cases, and p53 accumulation was present in 8%. Expression of six of the seven studied proteins showed no significant difference between mild, moderate, or florid ductal hyperplasias, indicating that there are no important cell biological differences with regard to the studied proteins between the lesions within this morphologically continuous spectrum. In addition, there were no differences between lesions with and without an invasive component. Cyclin D1 positivity was exclusively seen in lesions with 75% or more p27-positive nuclei. No significant correlations were found between other proteins. Twenty-three of 91 lesions (25%) had multiple events, of which five showed altered expressions of three or four proteins. In conclusion, altered protein expression of several proliferation- and apoptosis-related genes that are known to be involved in invasive breast cancer also may be found in usual ductal hyperplastic lesions, including several lesions with multiple events. This implies that usual ductal hyperplastic lesions may be among the earliest lesions within the breast oncogenetic spectrum.

Topics: Apoptosis; Biomarkers, Tumor; Breast; Carcinoma, Ductal, Breast; Cell Cycle Proteins; Cell Division; Cyclin D1; Epithelial Cells; Female; Gene Products, rex; Humans; Hyperplasia; Immunoenzyme Techniques; Ki-67 Antigen; Oncogene Protein p21(ras); Proto-Oncogene Proteins c-bcl-2; Receptor, ErbB-2; Tumor Suppressor Protein p53

There is controversy concerning the prognosis of breast cancers arising in women carrying loss of function mutations in the breast cancer susceptibility genes BRCA1 and BRCA2. This study was carried out to assess the likely hormone dependence of this group of tumours in comparison with an age and grade matched group of control sporadic tumours. We used quantitative immunohistochemical analysis for the oestrogen receptor (ER), progesterone receptor (PgR), cyclin D1 and pS2 on sections of primary tumours and ductal carcinoma in situ (DCIS). Expression of PgR (P < 0.05) and cyclin D1 (P < 0.01) was low in the BRCA1- and BRCA2-associated cancers compared with sporadic cases. The low frequency of expression of ER (9/40), PgR (2/40) cyclin D1 (5/36) and pS2 (5/36) in the familial tumours indicates that the majority of such tumours will be oestrogen insensitive and unlikely to respond to hormonal manipulation even at the in situ stage in their evolution. The low level of PgR (2/40 cases) suggests that there may be some abnormality of transactivating function of the ER in these tumours.

Topics: Adult; BRCA2 Protein; Breast Neoplasms; Carcinoma, Ductal, Breast; Cyclin D1; Female; Genes, BRCA1; Genetic Predisposition to Disease; Humans; Immunohistochemistry; Middle Aged; Neoplasm Proteins; Proteins; Receptors, Estrogen; Receptors, Progesterone; Transcription Factors; Trefoil Factor-1; Tumor Suppressor Proteins

Ductal carcinoma in situ (DCIS) of the breast is a heterogeneous disease clinically and biologically. The few available studies of its natural history implicate DCIS as a non-obligate precursor for invasive carcinoma. We have used fluorescence in situ hybridization (FISH) to detect gene amplification of the cell cycle regulator gene CCND1 in 88 examples of formalin-fixed, paraffin-embedded DCIS. Expression of its protein product cyclin D1 was detected by immunohistochemistry. CCND1 was amplified in 18% of DCIS cases. High grade DCIS was more likely to show amplification than low grade DCIS (32% versus 8%; P = 0.08). Gene amplification was associated with cyclin D1 protein expression (P = 0.001), although cyclin D1 was detected in cases that did not demonstrate gene amplification. Overall, cyclin D1 protein was detected in 50% of DCIS cases. Although only 2 of 23 (8%) cases of low grade DCIS had CCND1 amplification, over 50% (13/23) of these cases expressed cyclin D1 protein. Low grade DCIS had a higher mean percentage of nuclei expressing cyclin D1 than did intermediate or high grade DCIS (P = 0.007). Mechanisms other than gene amplification may be responsible for increased cyclin D1 protein in DCIS, especially in low grade DCIS. Identifying mechanisms that control cell cycle progression in DCIS may yield clues to its biological behavior.

Topics: Analysis of Variance; Breast Neoplasms; Carcinoma in Situ; Carcinoma, Ductal, Breast; Cyclin D1; Cyclins; Gene Amplification; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; In Situ Hybridization, Fluorescence; Multicenter Studies as Topic; Oncogene Proteins

Cyclin D1 (CCND1) and retinoblastoma (Rb) genes are cell cycle regulators which are altered in some breast carcinomas. However, the possible cooperation between CCND1 and Rb, as well as the influence and coincidence of their abnormalities in the proliferative capacity of mammary carcinoma cells in vivo, is still unknown. In order to assess both the significance of the CCND1 gene and Rb alterations in breast carcinomas and their relationship with the proliferative capacity of the tumours and other clinico-pathological factors, CCND1 mRNA expression was studied in 46 cases of primary breast carcinomas and matched normal tissue, 45 of which were also studied immunohistochemically, Rb expression was analysed in the same cases by immunohistochemistry, whereas the proliferative activity of the carcinoma was evaluated by flow cytometry. CCND1 mRNA was overexpressed in 19 tumours (41 per cent). Sixteen cases showed diffuse immunohistochemical expression, ten carcinomas had few positive cells, and 19 were absolutely negative. CCND1 mRNA and protein overexpression was associated with oestrogen receptor (ER) expression by the tumour. Interestingly, lack of ER expression was associated with a decreased CCND1 mRNA signal in non-overexpressed tumours. No association was observed between CCND1 mRNA or protein overexpression and tumour proliferation or other clinico-pathological parameters. Loss of Rb expression was observed in 26 per cent of the tumours. This abnormality was significantly associated with increased mean S-phase (P = 0.017) and decreased CCND1 mRNA expression in non-overexpressed tumours, supporting in vivo the postulated regulatory loop between Rb and CCND1 in vitro. We conclude that CCND1 up-regulation is not associated with increased proliferative activity in breast carcinomas, whereas its expression might be regulated in vivo by hormones and Rb. Loss of Rb expression is significantly associated with an increased proliferation of tumour cells, suggesting an important role in the progression of a subset of breast carcinomas, regardless of CCND1 abnormalities.

Topics: Blotting, Northern; Breast Neoplasms; Carcinoma; Carcinoma, Ductal, Breast; Carcinoma, Lobular; Cyclin D1; Cyclins; Flow Cytometry; Gene Expression; Genes, Retinoblastoma; Humans; Immunohistochemistry; Oncogene Proteins; Receptors, Estrogen

Protein complexes composed of cyclins and cyclin-dependent kinases control the orderly progression of mammalian cells through the cell cycle. The p27(Kip1) protein belongs to a family of cyclin-dependent kinase-inhibitory proteins that are negative regulators of cell cycle progression and have been proposed as candidate tumor suppressor genes. However, the p27(Kip1) gene is only rarely mutated in human primary breast carcinomas and breast cancer cell lines. To further address the role of p27(Kip1) in the development of human tumors, we determined by Western blot analysis the levels of expression of the p27(Kip1) protein in a series of human cancer cell lines and found that this protein is expressed at high levels in many of these cell lines, even during exponential growth. The levels of p27(Kip1) were significantly associated with the levels of cyclins D1 and E. In contrast to the high level of p27(Kip1) in breast cancer cell lines, three cell lines established from normal mammary epithelium expressed low levels of this protein. Cell synchronization studies demonstrated deregulation of the expression of p27(Kip1) throughout the cell cycle in two breast cancer cell lines but normal regulation in a normal mammary epithelial cell line. Immunohistochemical studies on p27(Kip1) expression in 52 primary human breast cancers indicated that this protein was also expressed at relatively high levels in 44% of the tumor samples, but it was barely detectable or undetectable in the remaining 56% of the samples. Additional studies are required to determine why some breast cancer cells express relatively high levels of p27(Kip1) despite its known role as an inhibitor of cell cycle progression.

Topics: Biomarkers, Tumor; Breast Neoplasms; Carcinoma in Situ; Carcinoma, Ductal, Breast; Carcinoma, Lobular; Cell Cycle; Cell Cycle Proteins; Cell Division; Cell Line, Transformed; Cyclin D1; Cyclin E; Cyclin-Dependent Kinase Inhibitor p27; Female; Gene Expression Regulation, Neoplastic; Humans; Microtubule-Associated Proteins; Neoplasm Proteins; Tumor Cells, Cultured; Tumor Suppressor Proteins

Somatic mutations in the retinoblastoma-1 gene (RB1) and loss of RB1 protein function have been implicated in a number of human malignancies, but the role of RB1 gene and protein abnormalities in ductal pancreatic cancer (DPCA) is virtually unknown. We therefore analyzed expression of the RB1 protein immunohistochemically and/or by western blotting in a total of 54 sporadic and eight familial cases of archival and frozen DPCA and in 18 pancreatic carcinoma cell lines by using the antibodies RB-WL-1, 84-B3-1, and PMG3-245. Mutations in the RB1 promotor region and exons 13-21 of the RB1 gene were likewise examined by single-strand conformation polymorphism (SSCP) analyses and DNA sequencing of genomic DNA from 30 microdissected primary pancreatic tumors and the pancreatic carcinoma cell lines. Moreover, amplification and expression of a major regulatory component of RB1 function, cyclin D1, were assessed by southern and immunohistochemical analyses, respectively. The DPCAs were heterogeneous in both the intensity of RB1 nuclear staining and the percentage of immunoreactive cells. The tumors often had areas where RB1 staining was weak or absent adjacent to normal pancreatic tissue; however, only two of 32 archival cases and one of 30 frozen cases of DPCA completely lacked RB1 nuclear staining. Immunohistochemical and western blot analyses of 18 pancreatic carcinoma cell lines demonstrated the absence of RB1 expression in only two cell lines, Capan-1 and QGP-1. Analyses of the RB1 gene and promotor region by SSCP and DNA sequencing largely confirmed the immunochemical findings. Three of 30 primary carcinomas had abnormalities revealed by SSCP analyses. In one case a single base-pair deletion was confirmed in exon 18 and resulted in premature termination and the absence of detectable RB1 protein. A second case had TAC-->TTC missense mutation in exon 13. The third primary carcinoma could not be reliably sequenced because it had a low percentage of epithelial cells. The cyclin D1 gene was not amplified in any of the primary pancreatic tumors or cell lines examined. These immunochemical and molecular analyses of the RB1 tumor suppressor gene and cyclin D1 proto-oncogene in a large series of human pancreatic cancers and cell lines indicate that RB1 and cyclin D1 alterations occur during the development of some human DPCAs. Nevertheless, it is probable that alterations in cell-cycle regulation in DPCAs more frequently involve pathways other than those involving RB1

Topics: Base Sequence; Blotting, Southern; Blotting, Western; Carcinoma, Ductal, Breast; Cell Line; Cyclin D1; Cyclins; Genes, Retinoblastoma; Humans; Immunohistochemistry; Molecular Sequence Data; Oncogene Proteins; Pancreatic Neoplasms; Polymorphism, Single-Stranded Conformational; Proto-Oncogene Mas; Retinoblastoma Protein; Tumor Cells, Cultured

Human p21 (also known as WAF1, CIP1, or SDI1) is a dual inhibitor of cyclin dependent kinases (CDKs) and the replication factor PCNA, which plays a role as a downstream mediator of the cell-cycle arrest induced by the tumor suppressor p53. To determine whether inactivation of downstream targets of p53 might contribute to cellular transformation, we have examined the integrity of the p21 gene in 36 invasive ductal breast carcinomas. Direct sequence analysis of the polymerase chain reaction-amplified p21 gene revealed a C to T transition in codon 94 that caused the substitution of a tryptophan for an arginine in a tumor specimen. This mutation was not detected in normal DNA extracted from the same patient nor in a polymerase chain reaction-restriction fragment length polymorphism of 50 unrelated individuals, indicating that it corresponds to a tumor-specific alteration. Functional analysis of the p21(R94W) protein produced in different eukaryotic and prokaryotic expression systems revealed that this mutation impaired the ability of p21 to inhibit CDKs. By contrast, the R94W mutant was unaltered in its ability to promote cyclin-CDK association as well as in its ability to bind proliferating cell nuclear antigen, thus leaving its putative functions as kinase activator or as inhibitor of replicative DNA synthesis intact. On the basis of these functional analysis, we propose that the Arg residue at position 94 is important for the CDK inhibitory role of p21.

Topics: Arginine; Base Sequence; Breast Neoplasms; Carcinoma, Ductal, Breast; Cell Cycle; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p21; Cyclin-Dependent Kinases; Cyclins; DNA Primers; Female; Humans; Molecular Sequence Data; Oncogene Proteins; Point Mutation; Protein Binding; Structure-Activity Relationship; Tryptophan

Topics: Breast Neoplasms; Carcinogenicity Tests; Carcinoma, Ductal, Breast; Carcinoma, Intraductal, Noninfiltrating; Cyclin D1; Cyclins; Female; Fibrocystic Breast Disease; Gene Expression Regulation; Humans; Oncogene Proteins; Oncogenes; RNA, Messenger

The elucidation of molecular alterations that occur during human breast cancer progression may contribute to the development of preventative strategies. Using in situ hybridizations on a cohort of 94 biopsy lesions, quantitatively increased cyclin D mRNA expression levels were observed in only 18% of benign lesions, which confer no or slightly increased breast cancer risk, and 18% of premalignant atypical ductal hyperplasias, which confer a four to fivefold increase in breast cancer risk. The transition to carcinoma was accompanied by frequent cyclin D mRNA overexpression in 76% of low-grade ductal carcinomas in situ, 87% of higher grade comedo ductal carcinomas in situ and 83% of infiltrating ductal breast carcinomas. The data identify a molecular event that may separate benign and premalignant human breast lesions from any form of breast carcinoma.

Topics: Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Intraductal, Noninfiltrating; Cohort Studies; Cyclin D1; Cyclins; Female; Fibrocystic Breast Disease; Gene Expression; Humans; Neoplasm Invasiveness; Oncogene Proteins; Precancerous Conditions; RNA, Messenger