cyclin-d1 and Vulvar-Neoplasms

Epithelial-mesenchymal transition (EMT) is associated with increased metastatic spread and poor prognosis. Data on vulvar carcinoma are limited.. Thirty-two cases of squamous cell carcinoma of the vulva (16 with and 16 without inguinal lymph node metastases) and their lymph node deposits were evaluated for immunohistochemical expression of EMT markers (vimentin, cyclin D1, e-cadherin), p16, p53 and Ki-67. Results of EMT-immunostainings were compared to lymph node involvement and expression of p53 and p16. The micro-anatomical staining pattern for EMT markers comparing the tumor center with the front of invasion was analysed in each tumor.. There was no difference in the expression of EMT markers between node negative and node positive tumors. Staining for vimentin and cyclin D1 was seen within tumor cells at the front of invasion in 100 and 84.4% of the tumors, respectively. The majority of cases (68.7%) showed negative or reduced staining for e-cadherin in this micro-anatomical localization. Tumor cells within the lymph node metastases showed positive staining for e-cadherin in 75% and for cyclin D1 in 49% of the cells but were negative for vimentin in 13 out of 16 cases (81.3%). Tumors with aberrant p53 staining represented a non-significant higher vimentin but significantly higher cyclin D1 expression at the front of invasion than those with p53 wild-type pattern.. The present study shows no differences in the expression of EMT markers between node positive and node negative vulvar cancers. The evaluation of immunostaining within the micro-anatomical context indicates that an EMT-phenotype is restricted to the tumor cells at the front of invasion. Paired analyses of vulvar carcinomas and their lymph node deposits suggest mesenchymal-epithelial transition (MET) in the metastatic deposits. Immunohistochemical staining results may suggest that EMT is more prevalent in vulvar cancer with aberrant p53 staining.

Topics: Biomarkers, Tumor; Cadherins; Cyclin D1; Epithelial-Mesenchymal Transition; Female; Humans; Lymph Nodes; Lymphatic Metastasis; Tumor Suppressor Protein p53; Vimentin; Vulvar Neoplasms

The aim of the present study was to screen differentially expressed miRNAs in vulvar squamous cell carcinoma (VSCC), observe the role of microRNA‑4712‑5p in VSCC and investigate its targets and regulatory mechanism. Differentially expressed miRNAs in human VSCC tissues were screened. microRNA‑4712‑5p was selected and its expression level was verified in clinical tissue samples and the VSCC cell line A431 by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) analysis. The overexpression vector of microRNA‑4712‑5p was prepared and transfected into A431 cells; subsequently, cell invasion and metastasis were examined by Cell Counting Kit‑8 and Transwell migration assays. Furthermore, the target gene of miRNA‑4712‑5p was predicted by bioinformatics and verified by The Dual‑Luciferase® Reporter (DLR™) Assay System. The expression of phosphatase and tensin homologue (PTEN) and its downstream proteins, such as protein kinase B (PKB; AKT), glycogen synthase kinase (GSK)3β and cyclin D1, were detected by western blot assays. The expression level of microRNA‑4712‑5p in VSCC tissues and the A431 cell line was found to be significantly increased, promoting proliferation and invasion of VSCC. The DLR™ assay indicated that PTEN was a target of miR‑4712‑5p. RT‑qPCR revealed that PTEN expression was markedly lower in VSCC tissues compared with that in adjacent tissues. After A431 cells were transfected with the miRNA‑4712‑5p overexpression vector, phospho‑AKT (p‑AKT) and cyclin D1 expression were notably increased, but miRNA‑4712‑5p‑targeted PTEN and phospho‑GSK3β (p‑GSK3β) protein markedly decreased. Therefore, microRNA‑4712‑5p can reduce the expression of PTEN, further affecting its downstream p‑AKT, p‑GSK3β and cyclin D1 signaling pathways, promoting the proliferation and invasion of VSCC.

Topics: 3' Untranslated Regions; Aged, 80 and over; Animals; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Proliferation; Cyclin D1; Female; Gene Expression Regulation, Neoplastic; Glycogen Synthase Kinase 3 beta; Humans; Mice; MicroRNAs; Middle Aged; Neoplasm Transplantation; Proto-Oncogene Proteins c-akt; PTEN Phosphohydrolase; Signal Transduction; Vulvar Neoplasms

To investigate cyclin D1 and fascin immunoreactivity in normal, reactive and neoplastic vulvar skin correlating the findings with p16 protein and Ki67 expression.. 66 vulvar biopsy or resection specimens demonstrating normal appearances, reactive epidermal changes, usual-type vulvar intraepithelial neoplasia (uVIN), differentiated-type VIN (dVIN), p16-positive squamous cell carcinoma (SCC) and p16-negative SCC were examined immunohistochemically for cyclin D1, fascin, Ki67 and p16 protein. Where applicable, expression patterns were compared in microanatomically distinct areas, particularly at the invasive front (deep tumour margin) of SCC.. Normal epidermis showed parabasal Ki67 and cyclin D1 staining while fascin labelled cells in the lower one-third of the epithelium. Reactive and dVIN specimens demonstrated mildly increased Ki67 and cyclin D1 expression that maintained parabasal polarity, whereas uVIN and p16-positive SCC were characterised by loss of cyclin D1 staining. However, in 14 of 20 p16-positive SCC small infiltrative tumour groups and single infiltrating cells at the invasive front showed a cyclin D1-positive/ Ki67-negative phenotype. In contrast, p16-negative SCC generally showed diffuse and concordant cyclin D1 and Ki67 labelling, including at the invasive margin. Fascin expression was increased in all VIN and SCC lesions.. Variations in cyclin D1 and Ki67 expression between p16-positive and p16-negative vulvar SCCs suggest different mechanisms of invasion in these tumour subgroups. Fascin is upregulated in vulvar squamous neoplasia but immunostaining does not discriminate in situ from invasive lesions nor putative human papilloma virus (HPV)-associated and HPV-independent SCCs.

Topics: Biomarkers, Tumor; Biopsy; Carcinoma in Situ; Carcinoma, Squamous Cell; Carrier Proteins; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Epidermis; Female; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Ki-67 Antigen; Microfilament Proteins; Up-Regulation; Vulva; Vulvar Neoplasms

Cyclin D1 (CCND1) belongs to the family of D-type cyclins involved in cell cycle progression, transcriptional regulation, and cell migration. CCND1 was found to be amplified and overexpressed in a variety of cancers, including some vulvar carcinoma cell lines. To determine the relationship of CCND1 copy number changes and CCND1 protein expression with clinicopathologic features and prognosis, 183 vulvar carcinomas were analyzed on a tissue microarray. Amplification was observed in 32 (22.4%) vulvar cancer specimens and was statistically related to the presence of regional lymph node metastases (P < .001). Detectable CCND1 expression was found in 139 (83.2%) of vulvar carcinomas, and 76 (45.5%) exhibited a moderate or strong expression. Increased levels of CCND1 expression were significantly related to higher patient age (P = .013), positive pN category (P = .004), and negative human papillomavirus status (P < .001). Basaloid as well as verrucous, warty-type, and mixed vulvar carcinomas showed lower CCND1 expression levels than keratinizing or nonkeratinizing tumors (P < .001 and P = .032, respectively). Elevated CCND1 expression levels and amplification of the CCND1 gene were closely connected in the present analysis (P < .001). Patient prognosis was independent from CCND1 amplification status and expression level (P = .57 each). In conclusion, CCND1 is amplified and overexpressed in a substantial proportion of vulvar carcinomas and associated with the occurrence of locoregional lymph node metastases, especially in human papillomavirus-negative tumors.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma; Cyclin D1; Female; Gene Amplification; Humans; Lymphatic Metastasis; Middle Aged; Prognosis; Vulva; Vulvar Neoplasms

Pilomatrix carcinoma, a malignant counterpart of pilomatrixoma, is a rare skin neoplasm composed of basaloid and shadow cells that characterize differentiation toward the hair matrix. The authors present a case of pilomatrix carcinoma of the clitoris, a very unusual location not previously reported. Diagnostic criteria and differential diagnoses are discussed. Pilomatrix carcinoma should be differentiated from benign pilomatrixoma and other carcinomas with shadow cells, including basal cell carcinoma with matrical differentiation and metastases of visceral carcinomas with shadow cells.

Topics: beta Catenin; Biomarkers, Tumor; Carcinoma, Basal Cell; Cell Nucleus; Clitoris; Cyclin D1; Diagnosis, Differential; Fatal Outcome; Female; Hair Diseases; Humans; Pilomatrixoma; Skin Neoplasms; Vulvar Neoplasms

Loss of retinoblastoma protein expression and overexpression of cyclin D1 have been implicated in the development and progression of some cancers. Paget's disease of the vulva (PDV) and Paget's disease of the breast (PDB) are uncommon conditions and the pathogenesis of these diseases is still unclear. The aim was to examine the expression of the retinoblastoma and cyclin D1 proteins in PDV and PDB and to correlate any differences between PDV and PDB, and in the presence or absence of an underlying carcinoma.. Seventy-two archival cases of PDV including 10 with invasive disease and 36 cases of PDB were evaluated immunohistochemically for the expression of cyclin D1 and retinoblastoma protein. Forty-four percent (32/72) of cases of PDV showed loss of expression of the retinoblastoma protein, compared with 67% (24/36) of PDB cases. Fifty-nine percent (41/69) of PDV overexpressed cyclin D1. In PDB, 8% (3/34) overexpressed cyclin D1. There were no significant differences in the expression of retinoblastoma and cyclin D1 in PDV cases with or without underlying invasive disease. There were significant differences between the expression of retinoblastoma (P = 0.03) and cyclin D1 (P < 0.001) in PDV compared with PDB.. The differences in the expression of cyclin D1 and retinoblastoma may indicate the differences in the pathogenesis of PDV and PDB.

Topics: Breast Neoplasms; Chi-Square Distribution; Cyclin D1; Female; Humans; Immunohistochemistry; Paget Disease, Extramammary; Paget's Disease, Mammary; Retinoblastoma Protein; Vulvar Neoplasms

The treatment of vulvar squamous cell carcinoma patients is often mutilating. Effort is being made to individualize treatment in order to reduce negative side effects for patients with good prognosis. Molecular markers have been able to predict patient outcome in several tumors. The aim of this study was to characterize the expression of cyclins D1, D3, E, and A in a comparatively large series of patients with vulvar squamous cell carcinoma and look for prognostic impact.. A total of 224 vulvar squamous cell carcinomas were immunohistochemically investigated for expression of cyclins D1, D3, E, and A using the biotin-streptavidin-peroxidase method and the OptiMax Plus automated cell staining system.. High protein levels of cyclin D1 (any positive nuclei) were found in 58 (26%) cases, cyclin D3 (> or =50% positive nuclei) in 61 (27%) cases, cyclin E (> or =50% positive nuclei) in 41 (18%) cases, and cyclin A (> or =5% positive nuclei) in 156 (70%) cases. No prognostic impact was found for the cyclins D1, D3, E, or A.. The high number of cases showing increased levels of cyclin A suggests that this protein may be important in the pathogenesis of vulvar squamous cell carcinoma. No prognostic impact was found for the cyclins D1, D3, E, or A.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cyclin A; Cyclin D1; Cyclin D3; Cyclin E; Cyclins; Female; Humans; Immunohistochemistry; Middle Aged; Predictive Value of Tests; Retrospective Studies; Vulvar Neoplasms

Vulval intraepithelial neoplasia (VIN) is defined histopathologically by distinctive abnormalities of cellular maturation and differentiation. To investigate the functional properties of VIN, the expression of several proteins involved in the regulation of the cell cycle as well as in situ DNA replication competence was analysed by immunohistochemistry. Snap-frozen vulval biopsies were graded as normal squamous epithelium (n=6), undifferentiated HPV positive VIN 1 (n=3), VIN 2 (n=8) and VIN 3 (n=20). Immunohistochemistry was performed using the following markers: cyclin D1 (expressed in middle/late G1), cyclin B1 (expressed in G2/early M), phosphorylated histone H3 (expressed during mitosis) and minichromosome maintenance (Mcm) proteins 2 and 5 (expressed during the cell cycle, but not in differentiated or quiescent cells). In situ DNA replication competence was used to identify S-phase cells. The percentage of positively stained nuclei in three representative microscopic fields was calculated per biopsy. In normal vulva, the expression of all markers was restricted to the proliferative compartment of the basal layer of the epithelium. In contrast in high-grade VIN, the majority of epithelial cells expressed the Mcm proteins from basal to superficial layer. The detection of cyclins B1 and D1, phospho-histone H3 and in situ DNA replication was also found through the full thickness of these lesions but by a lower proportion of the cells. This is consistent with these markers providing a series of 'snapshots' of the cell cycle status of individual cells. The low-grade VIN showed reduced expression of the cell cycle markers in relation to the level of dysplasia. The combination of these analyses establishes that the majority of VIN cells remain in a functional replicative or prereplicative state of the cell cycle. Clinical application of these analyses may provide a basis for improved diagnosis of VIN.

Topics: Biomarkers, Tumor; Biopsy; Carcinoma in Situ; Cell Cycle; Cell Cycle Proteins; Cyclin B; Cyclin B1; Cyclin D1; DNA Replication; DNA-Binding Proteins; DNA, Neoplasm; Female; Histones; Humans; Immunoenzyme Techniques; Minichromosome Maintenance Complex Component 2; Nuclear Proteins; Schizosaccharomyces pombe Proteins; Vulvar Neoplasms

Three different alterations in the p16/pRb/cyclin-D1 pathway (p16(INK4a)-promoter hypermethylation and expression of pRb and cyclin-D1) were investigated in a series of 38 cases of vulvar carcinoma (VC), 13 cases of vulvar intraepithelial neoplasia (VIN), and 21 cases of lichen sclerosus (LS). Paraffin blocks from 72 patients were selected for investigation of DNA methylation patterns in the CpG island of p16(INK4a) by methylation-specific polymerase chain reaction. Immunohistochemical studies for pRb and cyclin-D1 were performed using the standard avidin-biotin-peroxidase complex method. Epigenetic silencing of p16(INK4a) was detected in 68% of VC, 69.2% of VIN, and 42.8% of LS cases. Lack of pRb protein was found in 21% of VC, 0% of VIN, and 0% of LS cases. Overexpression of cyclin-D1 was found in 21% of VC, 30.8% of VIN, and 0% of LS cases. We conclude (1) that p16(INK4a) epigenetic inactivation most likely represents an early event, insufficient for malignant transformation, that may occur in clinically benign lesions such as LS; (2) that lack of pRb was only detected in fewer than one quarter of the carcinomas and could be considered a late secondary event; and (3) that cyclin-D1, which was overexpressed in VC and VIN, could contribute to the malignant transformation in association with p16 hypermethylation.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma; Carcinoma in Situ; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; DNA Primers; DNA, Neoplasm; Female; Humans; Immunoenzyme Techniques; Lichen Sclerosus et Atrophicus; Methylation; Middle Aged; Neoplasm Staging; Polymerase Chain Reaction; Retinoblastoma Protein; Vulvar Neoplasms

Cyclin D1 plays an essential regulatory role in the G1 phase of the cell cycle. The cyclin D1 gene is amplified in 20-50% of squamous cell carcinomas (SCCs), and the protein is overexpressed in up to 80% of SCCs. Our hypothesis was that gene transduction of antisense (AS) cyclin D1 in human SCCs in vivo would result in tumor reduction. A cyclin D1 cDNA was inserted into an E1/E3-deficient serotype 5 adenovirus (AS cyclin D1) in an AS orientation using homologous recombination. AS cyclin D1 transduction suppressed cyclin D1 protein expression in both cultured cells and tumors. AS cyclin D1 significantly inhibited cell proliferation by both [3H]thymidine incorporation in six SCC cell lines (P = 0.01-0.001) and the conversion of tetrazolium salt to formazan in four SCC cell lines (P = 0.01-0.004). Apoptosis detected in >25% of cells in each cell line 48 h after AS cyclin D1 transduction paralleled the reduction in cyclin D1 protein. Preformed SCCs transduced with AS cyclin D1 were significantly inhibited (P = 0.002-0.005), and apoptosis was prominent in the AS cyclin D1-treated tumors, but not in tumors treated with the control vector. These data extend prior in vitro and ex vivo results and indicate that AS cyclin D1 suppresses SCC growth both in vitro and in vivo through suppression of cyclin D1 protein expression, leading to cellular apoptosis. Our findings suggest that cyclin D1 may have a role in cell survival and that cyclin D1 AS therapy may be useful as an adjunct to standard treatment for SCC.

Topics: Adenoviruses, Human; Apoptosis; Carcinoma, Squamous Cell; Cell Cycle; Cell Division; Cyclin D1; DNA, Antisense; Female; Gene Amplification; Genetic Vectors; Head and Neck Neoplasms; Humans; Open Reading Frames; Skin Neoplasms; Tumor Cells, Cultured; Uterine Cervical Neoplasms; Vulvar Neoplasms

CYCLIN D1, a cell-cycle control gene, recently has been shown to be identical to an oncogene alternatively known as BCL-1 and PRAD1 and implicated in centrocytic lymphomas and parathyroid adenomas, respectively. PRAD1 complexes to the product of the retinoblastoma (Rb) tumor suppressor gene, an event followed by Rb inactivation. Squamous cell carcinomas of the cervix and vulva are gynecologic tumors in which human papillomaviruses have been implicated as an initiating event, and proteins derived from these viruses also complex with an inactivate Rb. Because of the overlap in some of the molecular processes mediated by human papillomaviruses and by the PRAD1 oncogene, the authors analyzed the PRAD1 (CYCLIN D1/BCL-1) genomic structure and expression in vulvar and cervical squamous cell carcinoma cell lines.. PRAD1 DNA and PRAD1 mRNA expression were assessed by Southern and Northern blotting, respectively, in 13 squamous cell carcinoma cell lines of gynecologic origin (10, cervical cancer; 3, vulvar cancer).. We found low baseline levels of a 4.5-kb PRAD1 transcript in a series of control cell lines, which were derived from normal fibroblasts, various hematologic malignancies, and a choriocarcinoma. PRAD1 mRNA overexpression (> or = 10-fold greater than that in control lines) was seen in all three vulvar carcinoma cell lines, two of which also showed amplification (5-fold and > 10-fold) of PRAD1 genomic sequences. Abnormalities of PRAD1 also were seen in 4 of the 10 cervical cancer cell lines and included overexpression of PRAD1 transcripts (3-9-fold) in 3 lines and rearrangement of PRAD1 DNA in an additional line that, however, did not shown any aberration in PRAD1 mRNA as discernible by Northern blotting. PRAD1 abnormalities were observed in three of the four cervical cell lines derived from metastatic sites and in one of the six cervical lines derived from primary tissue.. Seven of 13 squamous cell lines of gynecologic origin showed abnormalities of PRAD1. These abnormalities included amplification and rearrangement of DNA and overexpression of mRNA. The role of PRAD1 as a cell-cycle regulatory gene and its interactions with the Rb tumor suppressor gene suggests that PRAD1 deregulation may be a significant molecular event in the evolution of these tumors.

Topics: Blotting, Northern; Blotting, Southern; Carcinoma, Squamous Cell; Cyclin D1; Cyclins; Female; Gene Amplification; Gene Rearrangement; Humans; Oncogene Proteins; RNA, Messenger; Tumor Cells, Cultured; Uterine Cervical Neoplasms; Vulvar Neoplasms