cyclin-d1 and Papillomavirus-Infections

The incidence of HPV-positive oropharyngeal squamous cell carcinoma (OPSCC), which is both biologically and clinically distinct from tobacco- and alcohol-related OPSCC, is dramatically increasing. The finding that individuals with HPV-positive local/regionally advanced OPSCC have a significantly better prognosis than their negative counterparts have led to efforts to de-escalate treatment in those patients to avoid serious side effects and to improve their long-term quality of life, while maintaining treatment efficacy. Identifying diagnostic tests that are able to distinguish cancers etiologically associated with HPV is thus becoming a pressing challenge for researchers. The purpose of this review is to provide an overview of the diagnostic tools presently available to evaluate HPV status in patients with OPSCC and, in particular, to discuss their strengths and weaknesses in identifying those infections that are the real driving force in the oropharyngeal carcinogenesis process.

Topics: Alcohol Drinking; Antibodies, Viral; Biomarkers; Carcinoma, Squamous Cell; Cell Transformation, Viral; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Diagnosis, Differential; DNA, Neoplasm; DNA, Viral; Humans; Immunohistochemistry; In Situ Hybridization; Neoplasm Proteins; Oncogene Proteins, Viral; Oncogenes; Oropharyngeal Neoplasms; Papillomaviridae; Papillomavirus E7 Proteins; Papillomavirus Infections; Polymerase Chain Reaction; Repressor Proteins; Retinoblastoma Protein; Smoking; Tumor Suppressor Protein p53

Head and neck squamous cell carcinoma (HNSCC) is a common malignancy that continues to be difficult to treat and cure. In many organ systems and tumor types, there have been significant advances in the understanding of the molecular basis for tumorigenesis, disease progression and genetic implications for therapeutics. Although tumorigenesis pathways and the molecular etiologies of HNSCC have been extensively studied, there are still very few diagnostic clinical applications used in practice today. This review discusses current clinically applicable molecular markers, including viral detection of Epstein-Barr virus and human papillomavirus, and molecular targets that are used in diagnosis and management of HNSCC. The common oncogenes EGFR, RAS, CCND1, BRAF, and PIK3CA and tumor suppressor genes p53, CDKN2A and NOTCH are discussed for their associations with HNSCC. Discussion of markers with potential future applications is also included, with a focus on molecular alterations associated with targeted therapy resistance.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Class I Phosphatidylinositol 3-Kinases; Cyclin D1; Epstein-Barr Virus Infections; Gene Expression Regulation, Neoplastic; Genes, erbB-1; Genes, p16; Genes, p53; Genes, ras; Head and Neck Neoplasms; Humans; Papillomavirus Infections; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins B-raf; Receptors, Notch; Squamous Cell Carcinoma of Head and Neck

We aimed to establish image recognition and survival prediction models using a novel scoring system of cyclin D1 expression pattern in patients with human papillomavirus-negative oral or oropharyngeal squamous cell carcinoma.. The clinicopathological data of 610 patients with human papillomavirus-negative oral/oropharyngeal squamous cell carcinoma were analyzed retrospectively. Cox univariate and multivariate risk regression analyses were performed to compare cyclin D1 expression pattern scoring with the traditional scoring method-cyclin D1 expression level scoring-in relation to patients' overall and progression-free survival. An image recognition model employing the cyclin D1 expression pattern scoring system was established by YOLOv5 algorithms. From this model, two independent survival prediction models were established using the DeepHit and DeepSurv algorithms.. Cyclin D1 had three expression patterns in oral and oropharyngeal squamous cell carcinoma cancer nests. Superior to cyclin D1 expression level scoring, cyclin D1 expression pattern scoring was significantly correlated with the prognosis of patients with oral squamous cell carcinoma (p < 0.0001) and oropharyngeal squamous cell carcinoma (p < 0.05). Moreover, it was an independent prognostic risk factor in both oral squamous cell carcinoma (p < 0.0001) and oropharyngeal squamous cell carcinoma (p < 0.05). The cyclin D1 expression pattern-derived image recognition model showed an average test set accuracy of 78.48% ± 4.31%. In the overall survival prediction models, the average concordance indices of the test sets established by DeepSurv and DeepHit were 0.71 ± 0.02 and 0.70 ± 0.01, respectively.. Combined with the image recognition model of the cyclin D1 expression pattern, the survival prediction model had a relatively good prediction effect on the overall survival prognosis of patients with human papillomavirus-negative oral or oropharyngeal squamous cell carcinoma.

Topics: Carcinoma, Squamous Cell; Cyclin D1; Deep Learning; Head and Neck Neoplasms; Humans; Mouth Neoplasms; Oropharyngeal Neoplasms; Papillomavirus Infections; Prognosis; Retrospective Studies; Squamous Cell Carcinoma of Head and Neck

Dysregulation of fibroblast growth factor receptor (FGFR) signaling pathway has been observed in head and neck squamous cell carcinoma (HNSCC) and is a promising therapeutic target for selective tyrosine kinase inhibitors (TKIs). Potential predictive biomarkers for response to FGFR-targeted therapies are urgently needed. Understanding the epigenetic regulation of FGF pathway related genes, i.e. FGFRs, FGFs, and CCND1, could enlighten the way towards biomarker-selected FGFR-targeted therapies.. We performed DNA methylation analysis of the encoding genes FGFR1, FGFR2, FGFR3, FGFR4, FGF1-14, FGF16-23, and CCND1 at single CpG site resolution (840 CpG sites) employing The Cancer Genome Research Atlas (TCGA) HNSCC cohort comprising N = 530 tumor tissue and N = 50 normal adjacent tissue samples. We correlated DNA methylation to mRNA expression with regard to human papilloma virus (HPV) and gene amplification status. Moreover, we investigated the correlation of methylation with sensitivity to the selective FGFR inhibitors PD 173074 and AZD4547 in N = 40 HPV(-) HNSCC cell lines.. We found sequence-contextually nuanced CpG methylation patterns in concordance with epigenetically regulated genes. High methylation levels were predominantly found in the promoter flank and gene body region, while low methylation levels were present in the central promoter region for most of the analyzed CpG sites. FGFRs, FGFs, and CCND1 methylation differed significantly between tumor and normal adjacent tissue and was associated with HPV and gene amplification status. CCND1 promoter methylation correlated with CCND1 amplification. For most of the analyzed CpG sites, methylation levels correlated to mRNA expression in tumor tissue. Furthermore, we found significant correlations of DNA methylation of specific CpG sites with response to the FGFR1/3-selective inhibitors PD 173074 and AZD4547, predominantly within the transcription start site of CCND1.. Our results suggest an epigenetic regulation of CCND1, FGFRs, and FGFs via DNA methylation in HNSCC and warrants further investigation of DNA methylation as a potential predictive biomarker for response to selective FGFR inhibitors in clinical trials.

Topics: Carcinoma, Squamous Cell; Cohort Studies; Cyclin D1; DNA Methylation; Fibroblast Growth Factors; Head and Neck Neoplasms; Humans; Papillomavirus Infections; Protein-Tyrosine Kinases; Receptors, Fibroblast Growth Factor; Statistics, Nonparametric

The objective of the study is to investigate the biomarkers for diagnosis and prevention of human papillomavirus (HPV) infection-induced cervical cancer.. Cervical cancer tissues were collected from patients with cervical cancer, while noncancer tissues were collected from patients diagnosed with cervical lesions or uterine fibroids at the Chinese PLA General Hospital 301 and 309, China from December 2017 to June 2018. The cancer tissues were collected from the site of lesion, while the noncancer tissues were collected from similar anatomical locations. Quantitative real-time PCR, Western blot (WB), and immunohistochemistry (IHC) were used to detect the mRNA and protein levels of HPV E6/E7, RPRD1B (regulation of nuclear pre-mRNA domain containing 1B), cyclin D1, and transcription factor 4 (TCF4) between cervical cancer tissues and noncancer tissues. The correlation of HPV E6/E7, RPRD1B, cyclin D1, and TCF4 expressions was analyzed.. Twenty patients with cervical cancer and 27 controls without cervical cancer were included in this study. The mRNA expression of HPV E6/E7and RPRD1B was significantly higher in patients with cervical cancer than controls, while cyclin D1 mRNA expression was significantly lower in patients with cervical carcinoma in situ stage, compared with controls. RPRD1B protein expression was significantly higher in patients compared to controls when analyzed by IHC. TCF4 was significantly lower in clinical stage I and Ib of cervical cancer when analyzed by WB. The mRNA and protein expressions of RPRD1B and cyclin D1 were significantly different between patients younger than 50 years old, compared to patients 50 years and older.. HPV E6/E7 expression was associated with RPRD1B level in cervical cancer. The expression of RPRD1B and cyclin D1 in patients with cervical cancer might be affected by age.

Topics: Adult; Aged; Alphapapillomavirus; Case-Control Studies; Cell Cycle Proteins; China; Cyclin D1; Female; Humans; Middle Aged; Neoplasm Proteins; Oncogene Proteins, Viral; Papillomavirus E7 Proteins; Papillomavirus Infections; Repressor Proteins; RNA, Messenger; Transcription Factor 4; Uterine Cervical Neoplasms

Equine penile squamous cell carcinoma (EpSCC) is a relatively common cutaneous neoplasm with a poor prognosis. In this study, we aimed to determine the protein expression and colocalisation of FRA1, c-Myc, Cyclin D1, and MMP7 in normal (NT), tumour (T), hyperplastic epidermis and/or squamous papilloma (Hyp/Pap), poorly-differentiated (PDSCC), or well-differentiated (WDSCC) EpSCC using a tissue array approach. Further objectives were to correlate protein expression to (i) levels of inflammation, using a convolutional neural network (ii) equine papillomavirus 2 (EcPV2) infection, detected using PCR amplification. We found an increase in expression of FRA1 in EpSCC compared to NT samples. c-Myc expression was higher in Hyp/Pap and WDSCC but not PDSCC whereas MMP7 was reduced in WDSCC compared with NT. There was a significant increase in the global intersection coefficient (GIC) of FRA1 with MMP7, c-Myc, and Cyclin D1 in EpSCC. Conversely, GIC for MMP7 with c-Myc was reduced in EpSCC tissue. Inflammation was positively associated with EcPV2 infection in both NT and EpSCC but not Hyp/Pap. Changes in protein expression could be correlated with EcPV2 for Cyclin D1 and c-Myc. Our results evaluate novel biomarkers of EpSCC and a putative correlation between the expression of biomarkers, EcPV2 infection and inflammation.

Topics: Animals; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cyclin D1; Gene Expression Regulation, Neoplastic; Horses; Male; Matrix Metalloproteinase 7; Papillomaviridae; Papillomavirus Infections; Penile Neoplasms; Protein Binding; Proto-Oncogene Proteins c-fos; Proto-Oncogene Proteins c-myc; ROC Curve; Tissue Array Analysis

Head and neck squamous cell carcinoma (HNSCC) is a deadly disease in which precision medicine needs to be incorporated. We aimed to implement next-generation sequencing (NGS) in determining actionable targets to guide appropriate molecular targeted therapy in HNSCC patients.. Ninety-three tumors and matched blood samples underwent targeted sequencing of 244 genes using the Illumina HiSeq 2500 platform with an average depth of coverage of greater than 1,000×. Clinicopathological data from patients were obtained from 17 centers in Korea, and were analyzed in correlation with NGS data.. Ninety-two of the 93 tumors were amenable to data analysis. TP53 was the most common mutation, occurring in 47 (51%) patients, followed by CDKN2A (n=23, 25%), CCND1 (n=22, 24%), and PIK3CA (n=19, 21%). The total mutational burden was similar between human papillomavirus (HPV)-negative vs. positive tumors, although TP53, CDKN2A and CCND1 gene alterations occurred more frequently in HPV-negative tumors. HPV-positive tumors were significantly associated with immune signature-related genes compared to HPV-negative tumors. Mutations of NOTCH1 (p=0.027), CDKN2A (p < 0.001), and TP53 (p=0.038) were significantly associated with poorer overall survival. FAT1 mutations were highly enriched in cisplatin responders, and potentially targetable alterations such as PIK3CA E545K and CDKN2A R58X were noted in 14 patients (15%).. We found several targetable genetic alterations, and our findings suggest that implementation of precision medicine in HNSCC is feasible. The predictive value of each targetable alteration should be assessed in a future umbrella trial using matched molecular targeted agents.

Topics: Adult; Aged; Aged, 80 and over; Cadherins; Class I Phosphatidylinositol 3-Kinases; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Feasibility Studies; Female; Genetic Predisposition to Disease; Head and Neck Neoplasms; High-Throughput Nucleotide Sequencing; Humans; Male; Middle Aged; Molecular Targeted Therapy; Mutation; Papillomaviridae; Papillomavirus Infections; Receptor, Notch1; Republic of Korea; Sequence Analysis, DNA; Squamous Cell Carcinoma of Head and Neck; Tumor Suppressor Protein p53

Adenocarcinoma (ADC) of the uterine cervix (UC) is a rare form of cervical cancer (CC) caused due to the infection of Human Papilloma Virus (HPV). Cyclin D1 is one of the downstream targets of aberrantly activated Notch signaling, contribute to the etiology of CC. However, little is known about the role of Cyclin D1 in the modulation of cervical ADC and is controversial. The purpose of this study is to determine the role of Cyclin D1 protein and to elucidate the combined analysis with Notch signaling proteins in HPV associated ADCs of CC. A total of 60 biopsy samples (40 normal and 20 ADCs of CC) were analyzed for the expression of Cyclin D1 in HPV associated ADCs via immunohistochemistry and by immunoblotting. HPV-16 positive ADC patients showed a strong association with the Cyclin D1 expression (p = 0.007). The significant mean difference (p = 0.0001) and the pairwise comparison between Cyclin D1/JAG1 (p = 0.0001), and Cyclin D1/Notch-3 (p = 0.0001) were observed. The above Notch signaling proteins showed their synergistic role in modulating Cyclin D1 which in-turn regulates HPV-16 associated ADC of the uterine cervix (UC), affecting women's global health.

Topics: Adenocarcinoma; Adult; Blotting, Western; Cervix Uteri; Cyclin D1; Female; Global Health; Human papillomavirus 16; Humans; Immunohistochemistry; Jagged-1 Protein; Papillomavirus Infections; Receptor, Notch3; Uterine Cervical Neoplasms

Human immunodeficiency virus-infected individuals (HIVIIs) have a higher incidence of head and neck squamous cell carcinoma (HNSCC), and clinical and histopathological differences have been observed in their tumors in comparison with those of HNSCC patients without a human immunodeficiency virus (HIV) infection. The reasons for these differences are not clear, and molecular differences between HIV-related HNSCC and non-HIV-related HNSCC may exist. This study compared the mutational patterns of HIV-related HNSCC and non-HIV-related HNSCC.. The DNA of 20 samples of HIV-related HNSCCs and 32 samples of non-HIV-related HNSCCs was sequenced. DNA libraries covering exons of 18 genes frequently mutated in HNSCC (AJUBA, CASP8, CCND1, CDKN2A, EGFR, FAT1, FBXW7, HLA-A, HRAS, KEAP1, NFE2L2, NOTCH1, NOTCH2, NSD1, PIK3CA, TGFBR2, TP53, and TP63) were prepared and sequenced on an Ion Personal Genome Machine sequencer. DNA sequencing data were analyzed with Ion Reporter software. The human papillomavirus (HPV) status of the tumor samples was assessed with in situ hybridization, the MassARRAY HPV multiplex polymerase chain reaction assay, and p16 immunostaining. Mutation calls were compared among the studied groups.. HIV-related HNSCC revealed a distinct pattern of mutations in comparison with non-HIV-related HNSCC. TP53 mutation frequencies were significantly lower in HIV-related HNSCC. Mutations in HIV+ patients tended to be TpC>T nucleotide changes for all mutated genes but especially for TP53.. HNSCC in HIVIIs presents a distinct pattern of genetic mutations, particularly in the TP53 gene. HIV-related HNSCC may have a distinct biology, and an effect of the HIV virus on the pathogenesis of these tumors should not be ruled out. Cancer 2018;124:84-94. © 2017 American Cancer Society.

Topics: Adult; Aged; Cadherins; Carcinoma, Squamous Cell; Case-Control Studies; Caspase 8; Class I Phosphatidylinositol 3-Kinases; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Cyclin-Dependent Kinase Inhibitor p18; ErbB Receptors; F-Box-WD Repeat-Containing Protein 7; Female; Head and Neck Neoplasms; Histone Methyltransferases; Histone-Lysine N-Methyltransferase; HIV Infections; HLA-A Antigens; Humans; In Situ Hybridization; Intracellular Signaling Peptides and Proteins; Kelch-Like ECH-Associated Protein 1; LIM Domain Proteins; Male; Middle Aged; NF-E2-Related Factor 2; Nuclear Proteins; Papillomaviridae; Papillomavirus Infections; Protein Serine-Threonine Kinases; Proto-Oncogene Proteins p21(ras); Receptor, Notch1; Receptor, Notch2; Receptor, Transforming Growth Factor-beta Type II; Receptors, Transforming Growth Factor beta; Squamous Cell Carcinoma of Head and Neck; Transcription Factors; Tumor Suppressor Protein p53; Tumor Suppressor Proteins

Adenoid cystic carcinoma (ACC) in the lacrimal gland is a rare malignancy. P16 is encoded by the. Twenty-one ACCs in the lacrimal gland and ten matched healthy lacrimal glands were studied. P16 was detected with immunohistochemistry (IHC), and HPV was detected with in situ hybridization (ISH) and PCR in all cases. Other cell cycle proteins were also detected with IHC, including cyclin D1 and Ki67. The methylation status of the p16 promoter was detected with methylation-specific PCR (MSP) to further investigate the regulation of p16 expression.. The expression rates of p16 (47.6%, 10/21), cyclin D1 (100%, 21/21), and Ki67 (52.4%, 11/21) were increased in ACCs compared to healthy lacrimal glands (negative). The results showed p16 expression was limited to the inner ductal epithelial cells in the majority of the tubular and cribriform patterns. In solid ACCs, p16 was uniformly positive. HPV was negative in all 21 cases with ISH and PCR. P16 overexpression was associated with cyclin D1 overexpression (p=0.013). Only 13 cases were tested successfully with MSP. The expression rate of p16 methylation was 23.1% (3/13) of the ACCs. Compared with primary ACCs, recurrent ACCs showed higher p16, cyclin D1, and Ki67 expression (p=0.011, p=0.026, p=0.049, respectively).. In summary, p16 overexpression was cell-type dependent in ACCs in the lacrimal gland, while HPV infection was negative. P16 overexpression was unrelated to HPV infection. The mechanism of p16 overexpression needs to be further investigated in ACCs in the lacrimal gland.

Topics: Adolescent; Adult; Aged; Carcinoma, Adenoid Cystic; Case-Control Studies; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Cyclin-Dependent Kinase Inhibitor p18; Eye Neoplasms; Female; Gene Expression Regulation, Neoplastic; Humans; Ki-67 Antigen; Lacrimal Apparatus; Male; Middle Aged; Papillomaviridae; Papillomavirus Infections; Survival Analysis

Human papillomavirus (HPV) infection is associated with several genetic alterations including oncogene amplification, leading to increased aggression of tumors. Recently, a relationship between HPV infection and oncogene amplification has been reported, but this finding remains controversial. This study therefore investigated relationships between HPV infection and amplification of genes in the epidermal growth factor receptor (EGFR) signaling cascade in oral squamous cell carcinoma (OSCC). Extracted DNA from 142 formalin-fixed paraffin-embedded (FFPE) OSCC tissues was performed to investigate the copy number of EGFR, KRAS, c-myc and cyclin D1 genes using real-time polymerase chain reaction (RT-PCR) and compared with calibrators. A tissue microarray of OSCC tissues was used for detection of c-Myc expression and HPV infection by immunohistochemistry and HPV E6/E7 RNA in situ hybridization, respectively. HPV infection was also investigated using PCR and RT-PCR. Of the 142 OSCC samples, 81 (57%) were HPV-infected cases. The most frequently amplified gene was c-myc (55.6%), followed by cyclin D1 (26.1%), EGFR (23.9%) and KRAS (19.7%). Amplification of c-myc was significantly associated with levels of its protein product. EGFR amplification was also significantly associated with amplification of genes in the signaling cascade: KRAS (50.0%), c-myc (34.2%) and cyclin D1 (46.0%). Interestingly, HPV infection was significantly associated with amplification of both EGFR (76.5%) and cyclin D1 (73.0%). Only cyclin D1 amplification was significantly associated with severity of OSCC histopathology. HPV infection may play an important synergistic role in amplification of genes in the EGFR signaling cascade, leading to increased aggression in oral malignancies.

Topics: Aged, 80 and over; Carcinoma, Squamous Cell; Cell Line; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; DNA, Viral; ErbB Receptors; Female; Gene Amplification; HeLa Cells; Humans; Male; Mouth Neoplasms; Papillomaviridae; Papillomavirus Infections; Proto-Oncogene Proteins c-myc

Head and neck squamous cell carcinoma (HNSCC) is the sixth most prevalent cancer worldwide with rates of HPV-positive oropharyngeal squamous cell carcinoma (OPSCC) dramatically increasing. The overexpression of enhancer of zeste homolog 2 (EZH2), a histone methyltransferase responsible for the trimethylation at lysine 27 of histone 3 (H3K27me3), is associated with a poor clinical prognosis and aggressive HPV-positive phenotypes.. We utilized three EZH2 pathway inhibitors, GSK-343, DZNeP, and EPZ-5687, and tested their efficacy in two HPV-positive and two HPV-negative OPSCC cell lines.. Treatment with GSK-343 decreased H3K27me3 in all cell lines and treatment with DZNeP decreased H3K27me3 in only HPV-negative cell lines as determined by Western blot. Cells treated with EPZ-5687 displayed no appreciable change in H3K27me3. Epigenetic effect on gene expression was measured via ddPCR utilizing 11 target probes. Cells treated with DZNeP showed the most dramatic expressional changes, with decreased EGFR in HPV-positive cell lines and an overall increase in proliferation markers in HPV-negative cell lines. GSK-343-treated cells displayed moderate expressional changes, with CCND1 increased in HPV-positive cell lines and decreased TP53 in HPV-negative SCC-1. EPZ-5687-treated cell lines displayed few expressional changes overall. Only DZNeP-treated cells displayed anti-proliferative characteristics shown in wound-healing assays.. Our findings suggest that EZH2 inhibitors are a viable therapeutic option for the role of epigenetic effect, potentially sensitizing tumors to current chemotherapies or limiting cell differentiation.

Topics: Carcinoma, Squamous Cell; Cell Line, Tumor; Cyclin D1; Enhancer of Zeste Homolog 2 Protein; Epigenesis, Genetic; ErbB Receptors; Gene Expression Regulation, Neoplastic; Histones; Humans; Indazoles; Methylation; Oropharyngeal Neoplasms; Papillomavirus Infections; Pyridones

While human papillomavirus (HPV) is an accepted risk factor for oropharyngeal squamous cell carcinoma (SCC), its aetiological role in oral cavity SCC remains unclear. This study aimed to determine the HPV prevalence in an Australian population.. DNA was extracted from 63 formalin-fixed paraffin-embedded tumour specimens histologically confirmed as SCC of the oral cavity, diagnosed during 2006-2012. Clinical data were extracted from medical records. HPV presence was determined by polymerase chain reaction. Positive samples were typed by sequencing. Immunohistochemistry was used to assess p16. Five of the 63 tumours (8%) were positive for HPV DNA (three HPV-16 positive and two HPV-18 positive). Two tumours overexpressed p16. This study has identified a low prevalence of high-risk HPV in Queensland, Australia.

Topics: Adult; Aged; Aged, 80 and over; Australia; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cyclin D1; Female; Human papillomavirus 16; Human papillomavirus 18; Human Papillomavirus DNA Tests; Humans; Male; Middle Aged; Mouth; Mouth Neoplasms; Oropharyngeal Neoplasms; Papillomaviridae; Papillomavirus Infections; Prevalence; Queensland; Risk Factors

Squamous cell carcinomas of the hypopharynx (HPSCC) and oropharynx (OPSCC) have markedly different patient outcomes. Differences in HPV prevalence between these two patient groups may account for some of this difference, but other molecular markers of prognosis or pathological phenotype have not been established. Copy number gain of oncogenes is a well-established molecular change contributing to HNSCC development. Quantitative PCR was used to explore copy number gains of specific genes (3q-PIK3CA, TP63; 11q13.3-CCND1, ANO1) in tumor DNA recovered from HPSCC (n = 48) and OPSCC (n = 52) patients. Associations between copy number gain, patient demographics, HPV/p16INK4a status and pathological stage were examined. HPV/p16 prevalence in HPSCC and OPSCC groups was 2.1% and 46.0%, respectively. HPSCCs had frequent gains of CCND1 (56.3%) and ANO1 (56.3%) but few gains of PIK3CA (6.3%). By contrast, OPSCCs had significantly fewer CCND1 (23.1%) and ANO1 (17.3%) gains, and significantly more PIK3CA (26.9%) gains. A mutually exclusive relationship between HPV/p16 and 11q13.3 gains was observed in OPSCCs, while PIK3CA and TP63 gains were similar across HPV-associated and smoking/alcohol-associated patients. ANO1 gain was significantly linked to tumor pathology in HPSCC, associating with nodal metastasis and smaller and less invasive tumors at presentation (P = 0.010). Our results provide a convincing link between a specific molecular change and disease phenotype that appears unique to our HPSCC population, supporting a model of 11q13.3 in promoting metastatic disease progression in HNSCC, and suggest a role for ANO1 as a molecular marker of metastatic disease. © 2016 Wiley Periodicals, Inc.

Topics: Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Chromosomes, Human, Pair 11; Class I Phosphatidylinositol 3-Kinases; Cyclin D1; DNA Copy Number Variations; Female; Follow-Up Studies; Humans; Hypopharyngeal Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Staging; Oropharyngeal Neoplasms; Papillomaviridae; Papillomavirus Infections; Phosphatidylinositol 3-Kinases; Prognosis; Survival Rate

The purpose of this study was to investigate roles of human papillomavirus (HPV) infection and stathmin in sinonasal inverted papilloma (SNIP).. HPV DNA detection was performed by the fluorescence-based polymerase chain reaction (PCR) method. Stathmin protein expression was investigated by the immunohistochemistry method and mRNA expression of stathmin, Kif2a, and cyclin D1 were assessed by real-time PCR in SNIP and control subjects.. The positive rate of HPV DNA detected in SNIP was about 53.6% (15 of 28). Recurrent cases showed a higher rate of HPV infection compared with initial cases and higher Krouse stage (T3 + T4) cases showed higher rate of HPV infection than lower Krouse stage (T1 + T2) cases. Stronger expression of stathmin, Kif2a, and cyclin D1 were observed in SNIP, especially HPV(+) SNIP.. HPV infection was closely associated with recurrence and progression of SNIP. Stathmin is a valuable prognostic marker and could be considered as a therapeutic target in patients with SNIP.

Topics: Adolescent; Adult; Aged; Biomarkers, Tumor; Case-Control Studies; Cyclin D1; DNA, Viral; Female; Humans; Immunohistochemistry; Kinesins; Male; Middle Aged; Nasal Mucosa; Neoplasm Recurrence, Local; Nose Neoplasms; Papilloma, Inverted; Papillomaviridae; Papillomavirus Infections; Paranasal Sinus Neoplasms; Polymerase Chain Reaction; RNA, Messenger; Stathmin; Up-Regulation; Young Adult

We conducted a large international study to estimate fractions of head and neck cancers (HNCs) attributable to human papillomavirus (HPV-AFs) using six HPV-related biomarkers of viral detection, transcription, and cellular transformation.. Formalin-fixed, paraffin-embedded cancer tissues of the oral cavity (OC), pharynx, and larynx were collected from pathology archives in 29 countries. All samples were subject to histopathological evaluation, DNA quality control, and HPV-DNA detection. Samples containing HPV-DNA were further subject to HPV E6*I mRNA detection and to p16(INK4a), pRb, p53, and Cyclin D1 immunohistochemistry. Final estimates of HPV-AFs were based on HPV-DNA, HPV E6*I mRNA, and/or p16(INK4a) results.. A total of 3680 samples yielded valid results: 1374 pharyngeal, 1264 OC, and 1042 laryngeal cancers. HPV-AF estimates based on positivity for HPV-DNA, and for either HPV E6*I mRNA or p16(INK4a), were 22.4%, 4.4%, and 3.5% for cancers of the oropharynx, OC, and larynx, respectively, and 18.5%, 3.0%, and 1.5% when requiring simultaneous positivity for all three markers. HPV16 was largely the most common type. Estimates of HPV-AF in the oropharynx were highest in South America, Central and Eastern Europe, and Northern Europe, and lowest in Southern Europe. Women showed higher HPV-AFs than men for cancers of the oropharynx in Europe and for the larynx in Central-South America.. HPV contribution to HNCs is substantial but highly heterogeneous by cancer site, region, and sex. This study, the largest exploring HPV attribution in HNCs, confirms the important role of HPVs in oropharyngeal cancer and drastically downplays the previously reported involvement of HPVs in the other HNCs.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cross-Sectional Studies; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; DNA, Viral; Female; Genotype; Head and Neck Neoplasms; Human papillomavirus 16; Humans; Immunohistochemistry; International Cooperation; Male; Middle Aged; Oropharyngeal Neoplasms; Papillomaviridae; Papillomavirus Infections; Predictive Value of Tests; Salivary Proline-Rich Proteins; Tumor Suppressor Protein p53

Persistent infection with oncogenic human papillomavirus viruses (HPVs) is a casual factor for cervical cancer and its precursors, and the abnormal constitutive expression of viral oncoprotein E6 is a key event during the malignant transformation. Here, we performed miRNA microarray to identify changes of miRNAs following ectopic HPV16 E6 overexpression in HEK293T cells and found miR-2861 was greatly decreased in both HEK293T and HaCaT cells expressing HPV16 E6 compared to vector control. Further, we demonstrated a biological link among HPV16 E6, miR-2861, EGFR, AKT2, and CCND1 in cervical cancer cells. We showed that miR-2861 was downregulated in cervical cancer tissues and negatively correlated with advanced tumor stage and lymph node metastasis. Overexpression of miR-2861 suppressed cervical cancer cell proliferation and invasion and enhanced apoptosis. Subsequent investigation revealed that EGFR, AKT2, and CCND1 were all the direct targets of miR-2861. Importantly, silencing EGFR, AKT2, and/or CCND1 recapitulated the cellular effects seen upon miR-2861 overexpression. Restoration of EGFR, AKT2, and/or CCND1 counteracted the effects of miR-2861 expression. Thus, we identified a new pathway employing miR-2861, EGFR, AKT2, and CCND1 that may mediate HPV16 E6 induced initiation and progression of cervical cancer.

Topics: Cell Line, Tumor; Cell Transformation, Viral; Cyclin D1; ErbB Receptors; Female; Gene Expression Regulation, Neoplastic; HEK293 Cells; Human papillomavirus 16; Humans; MicroRNAs; Neoplasm Metastasis; Neoplasm Staging; Oligonucleotide Array Sequence Analysis; Oncogene Proteins, Viral; Papillomavirus Infections; Proto-Oncogene Proteins c-akt; Repressor Proteins; Signal Transduction; Uterine Cervical Neoplasms

The role of human papillomavirus (HPV) in sinonasal inverted papillomas (IPs) is controversial. Determining the prevalence of HPV infection and its impact on the molecular biology of these tumors is critical to characterizing its role in the pathogenesis of IPs.. A total of 112 paraffin-embedded IPs from 90 patients were studied. A tissue microarray was constructed and stained for p16, p53, epidermal growth factor receptor (EGFR), and cyclin D1. HPV presence and types were determined using PGMY 09/11 primers and integration using HPV 11 detection of integrated papillomavirus sequences by ligation-mediated polymerase chain reaction (DIPS-PCR).. HPV was detected in 11 of 90 (12%) patients. HPV 11 was found in 9 samples. HPV 6 and HPV 27 were found in 1 sample each. EGFR staining proportion was higher in HPV-positive IPs vs HPV-negative specimens (56.2% vs 23.6%; p = 0.009). Differences in p16, p53, and cyclin D1 staining were not significant. HPV-positive lesions tend to progress to malignancy (p = 0.064). Three samples were analyzed for integration. Viral integration was found in both malignant tumors but not in the precursor IP.. Degradation of p53 and p16/cyclin D1 dysregulation are not important mechanisms in low-risk HPV-related IP. The low prevalence of HPV in this series indicates it is not a main etiological factor for IPs; however, when present, low-risk HPV may contribute to the biology of IPs through an increase of EGFR expression and a predisposition for malignant progression by integration into the cellular genome.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Disease Progression; DNA, Viral; ErbB Receptors; Female; Human papillomavirus 11; Humans; Immunohistochemistry; Male; Neoplasm Proteins; Neoplasm Recurrence, Local; Papilloma, Inverted; Papillomavirus Infections; Paranasal Sinus Neoplasms; Polymerase Chain Reaction; Tumor Suppressor Protein p53

Current conventional treatment modalities in head and neck squamous cell carcinoma (HNSCC) are nonselective and have shown to cause serious side effects. Unraveling the molecular profiles of head and neck cancer may enable promising clinical applications that pave the road for personalized cancer treatment. We examined copy number status in 36 common oncogenes and tumor suppressor genes in a cohort of 191 oropharyngeal squamous cell carcinomas (OPSCC) and 164 oral cavity squamous cell carcinomas (OSCC) using multiplex ligation probe amplification. Copy number status was correlated with human papillomavirus (HPV) status in OPSCC, with occult lymph node status in OSCC and with patient survival. The 11q13 region showed gain or amplifications in 59% of HPV-negative OPSCC, whereas this amplification was almost absent in HPV-positive OPSCC. Additionally, in clinically lymph node-negative OSCC (Stage I-II), gain of the 11q13 region was significantly correlated with occult lymph node metastases with a negative predictive value of 81%. Multivariate survival analysis revealed a significantly decreased disease-free survival in both HPV-negative and HPV-positive OPSCC with a gain of Wnt-induced secreted protein-1. Gain of CCND1 showed to be an independent predictor for worse survival in OSCC. These results show that copy number aberrations, mainly of the 11q13 region, may be important predictors and prognosticators which allow for stratifying patients for personalized treatment of HNSCC.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Squamous Cell; CCN Intercellular Signaling Proteins; Chromosome Aberrations; Chromosomes, Human, Pair 11; Cyclin D1; Disease-Free Survival; DNA Copy Number Variations; Female; Genes, Tumor Suppressor; Human papillomavirus 16; Human Papillomavirus DNA Tests; Humans; Lymphatic Metastasis; Male; Middle Aged; Mouth Neoplasms; Neoplasm Staging; Oncogenes; Oropharyngeal Neoplasms; Papillomavirus Infections; Precision Medicine; Proto-Oncogene Proteins; Young Adult

The p16INK4a protein is a principal cyclin-dependent kinase inhibitor that decelerates the cell cycle. Abnormally high levels of p16INK4a are commonly observed in human papillomavirus (HPV)-positive head and neck squamous cell carcinomas (HNSCC). We and others found that p16INK4a overexpression is associated with improved therapy response and survival of patients with HNSCC treated with radiotherapy. However, the functional role of p16INK4a in HNSCC remains unexplored. Our results implicate p16INK4a in regulation of homologous recombination-mediated DNA damage response independently from its role in control of the cell cycle. We found that expression of p16INK4a dramatically affects radiation sensitivity of HNSCC cells. p16INK4a overexpression impairs the recruitment of RAD51 to the site of DNA damage in HPV-positive cells by downregulating of cyclin D1 protein expression. Consistent with the in vitro findings, immunostaining of HNSCC patient samples revealed that high levels p16INK4a expression significantly correlated with decreased cyclin D1 expression. In summary, these findings reveal an unexpected function of p16INK4a in homologous recombination-mediated DNA repair response and imply p16INK4a status as an independent marker to predict response of patients with HNSCC to radiotherapy.

Topics: Carcinoma, Squamous Cell; Cell Line, Tumor; Chemoradiotherapy; Cyclin D1; Cyclin-Dependent Kinase 4; Cyclin-Dependent Kinase 6; Cyclin-Dependent Kinase Inhibitor p16; DNA Damage; Head and Neck Neoplasms; Humans; Kaplan-Meier Estimate; Papillomavirus Infections; Rad51 Recombinase; Radiation Tolerance; Recombinational DNA Repair; Treatment Outcome

Human papillomavirus-related oropharyngeal squamous cell carcinoma has a unique biology and improved prognosis. A new focus is to identify prognostic biomarkers specifically in this human papillomavirus-positive cohort. We analyzed cyclin D1 immunostaining on a tissue microarray of patients with known clinical follow-up and p16 and human papillomavirus status (by E6/E7 RNA in situ hybridization). Cyclin D1 staining was read visually and digitally. Cutoffs of 5%, 10%, and 30% were separately analyzed as was linear intensity data derived from the image analysis. For the 202 tumors, cyclin D1 expression was > 10% in 25.7% (visual) and 35.5% (digital) of the cases. It was > 30% in 15.8% (visual) and 16.5% (digital) of the cases. High cyclin D1 by both methods, cutoffs, and expression intensity was associated with poorer overall, disease-free, and disease-specific survival in univariate analysis. However, low cyclin D1 expression was also tightly associated with human papillomavirus RNA (P < 1.0 × 10(-18) for all cutoffs) and p16 positivity (P < 1.0 × 10(-14) for all cutoffs). In multivariate analysis using the digital 30% cutoff (the strongest cyclin D1 assessment method), only T stage, p16 status, smoking, and treatment approach associated with survival. Intensity of cyclin D1 expression did, however, significantly substratify the human papillomavirus RNA-positive patients into prognostic subgroups independent of other variables. In summary, cyclin D1 overexpression correlates strongly with patient survival in oropharyngeal squamous cell carcinoma, but its relationship with human papillomavirus status is very tight, and the complex nature of this correlation likely limits any clinical application for cyclin D1 assessment.

Topics: Biomarkers, Tumor; Carcinoma, Squamous Cell; Cyclin D1; Disease-Free Survival; Female; Humans; Immunohistochemistry; In Situ Hybridization; Kaplan-Meier Estimate; Male; Middle Aged; Oropharyngeal Neoplasms; Papillomavirus Infections; Prognosis; Proportional Hazards Models; Tissue Array Analysis

High-risk human papillomaviruses are closely associated with cervical cancer and its precursor lesions through interactions between the E6 and E7 oncoproteins and the cell-cycle regulatory proteins, such as p53 and pRb, respectively. As other molecules involved in the cell-cycle control seem to be important for human papillomavirus (HPV)-mediated cervical carcinogenesis, we have analyzed the expression of p53, p21, p16, cyclin D1, and Ki-67 and the presence of HPV (HPV pool and HPV-16) by immunohistochemical studies using tissue microarray in low squamous intraepithelial lesions (n=50), high squamous intraepithelial lesions (n=98), and cervical carcinoma (n=18). We have found a significant increase in the expression of p16 and p21 (P<0.001) from low- to high-grade lesions and cancer. In contrast, cyclin D1 expression showed a significant decrease in more severe lesions (P<0.001). p16, Ki-67, p21, and p53 positivity increased with the cell-layer level and the lesion severity, with stronger correlations being observed for p16 and Ki-67. High positivity for HPV pool (96.3%) and HPV-16 (77.5%) immunostaining was detected in all cases, with an association between p16 and cyclin D1 expression and HPV-16 infection. Our tissue microarray results corroborate the usefulness of the immunohistochemical assessment of cell-cycle biomarkers in distinguishing different groups of precursor lesions of the cervix and cervical carcinoma.

Topics: Biomarkers, Tumor; Brazil; Carcinogenesis; Carcinoma in Situ; Cell Cycle; Cervix Uteri; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Cyclin-Dependent Kinase Inhibitor p21; Female; Human papillomavirus 16; Humans; Immunohistochemistry; Ki-67 Antigen; Neoplasm Proteins; Papillomaviridae; Papillomavirus Infections; Precancerous Conditions; Tissue Array Analysis; Tumor Suppressor Protein p53; Uterine Cervical Neoplasms

Cervical cancer constitutes the second most common cancer in women. It is evident from earlier studies that epidermal growth factor (EGF) is a mitogen, in that it mimics the function of estrogen by mediating cross-talk with other oncoproteins. Although epidermal growth factor receptor (EGFR) is highly expressed in breast and ovarian tumor tissues, its regulation by the exogenous source of its ligand EGF in human papillomavirus (HPV)-associated cervical cancer remains unclear. In this study, we addressed the question of whether EGF is required for the proliferation of HPV-positive cervical cancer cells and what mechanisms are involved. To determine this, we conducted a series of studies using HPV-positive human cervical cancer cells, CaSki and HeLa, and stimulated the cells with EGF. Our findings suggest that 6 h of stimulation with 10 ng/ml of EGF is sufficient to induce cell cycle progression associated with a significant increase in DNA synthesis, EGFR, COX-2 and cyclin D1 levels. Consistently, cellular localization and Western blot analysis for p-EGFR (Try-1045) protein showed an increase after EGF stimulation. Using siRNA gene knockdown assays we have shown that cyclin D1 siRNA has a significant negative effect on EGFR and inhibit cell growth independent of COX-2 levels. In summary, our findings reveal that an exogenous EGF stimulation may enhance HPV-related cervical cancer cell proliferation by activating EGFR and cyclin D1 that is independent of COX-2 levels, suggesting that the inhibitors of EGFR and cyclin D1 may be effective against cervical cancer cell proliferation.

Topics: Cell Cycle; Cell Growth Processes; Cell Line, Tumor; Cyclin D1; Cyclooxygenase 2; Epidermal Growth Factor; ErbB Receptors; Female; HeLa Cells; Human papillomavirus 16; Human papillomavirus 18; Humans; Molecular Targeted Therapy; Papillomavirus Infections; Transcription, Genetic; Uterine Cervical Neoplasms

Human papillomavirus (HPV)-related oropharyngeal squamous cell carcinoma (SCC) exhibits distinct patterns worldwide, but its prevalence has not been extensively evaluated in Korea. The E7 oncogene-mediated carcinogenesis and its meaning are yet to be uncovered for oropharyngeal SCCs.. In a Korean oropharyngeal SCC cohort, epidemiological indicators, HPV, and G1 cell cycle marker expressions were correlated with survival.. Among 93 surgically treated patients with oropharyngeal SCCs, 49.5% were HPV+, which were significantly younger, and predominantly nonsmoking. They demonstrated better survival than HPV- (94% vs 60%). Patients who were HPV+ with oropharyngeal SCCs expressed higher p16, cyclin-dependent kinase 4 (cdk4), and lower pRb. The p16 (hazard ratio [HR] 2.39), pRb (HR 2.13), and CCND1 (HR 2.09) correlated with survival. Notably, combined markers like p16/cdk4 ratio (HR 2.47) and cdk4+CCND1 sum (HR 2.65) were more significantly correlated.. Incidence of HPV-related oropharyngeal SCC in Korea is similar to U.S.-European data. HPV presence correlates with improved survival. Expression ratios of G1 markers may predict survival of oropharyngeal SCCs better than each marker alone.

Topics: Adult; Aged; Aged, 80 and over; Analysis of Variance; Biomarkers, Tumor; Biopsy, Needle; Carcinoma, Squamous Cell; Cell Cycle; Cohort Studies; Comorbidity; Confidence Intervals; Cyclin D1; Cyclin-Dependent Kinase 4; Cyclin-Dependent Kinase Inhibitor p16; Disease-Free Survival; DNA, Viral; Female; Humans; Immunohistochemistry; Kaplan-Meier Estimate; Korea; Male; Middle Aged; Neoplasm Proteins; Neoplasm Recurrence, Local; Oropharyngeal Neoplasms; Papillomaviridae; Papillomavirus Infections; Polymerase Chain Reaction; Prevalence; Prognosis; Proportional Hazards Models; Regression Analysis; Retrospective Studies; Risk Assessment; Survival Analysis

There is increasing use of multiple molecular markers to predict prognosis in human cancer. Our aim was to examine the prognostic significance of cyclin D1 and retinoblastoma (pRb) expression in association with human papillomavirus (HPV) status in oropharyngeal squamous cell carcinoma. Clinical records and specimens of 226 patients with follow-up from 1 to 235 months postdiagnosis were retrieved. Tumor HPV status was determined by HPV E6-targeted multiplex real-time PCR/p16 semiquantitative immunohistochemistry and cyclin D1 and pRb expression by semiquantitative immunohistochemistry. Determinants of recurrence and mortality hazards were modeled using Cox regression with censoring at dates of last follow-up. The HPV-positivity rate was 37% (91% type 16). HPV was a predictor of recurrence, an event (recurrence or death) and death after adjustment for clinicopathological variables. There were inverse relationships between HPV status and cyclin D1 and pRb. On univariate analysis, cyclin D1 predicted locoregional recurrence, event and death and pRb predicted event and death. Within the HPV-positive group, after adjusting for clinicopathological factors, patients with cyclin D1-positive cancers had up to a eightfold increased risk of poor outcome relative to those with cyclin D1-negative tumors. However, within the HPV-negative group, there was only a very small adjusted increased risk. A combination of pRb and HPV did not provide additional prognostic information. Our data provide the first evidence that a combination of HPV and cyclin D1 provides more prognostic information in oropharyngeal cancer than HPV alone. If findings are confirmed, treatment based on HPV and cyclin D1 may improve outcomes.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cyclin D1; Female; Humans; Male; Middle Aged; Oropharyngeal Neoplasms; Papillomavirus Infections; Prognosis; Recurrence; Retinoblastoma Protein; Treatment Outcome

To evaluate the prognostic relevance of key cell cycle regulatory proteins p53, p16(INK4a), pRb and Cyclin D1 expression, the presence of high risk HPVs and their association with clinicopathological parameters and the clinical follow up in ovarian cancer patients.. 53 cases of primary ovarian serous carcinomas were immunohistochemically examined for the expression of p53, p16(INK4a), pRb and Cyclin D1 proteins. Tumor DNA was extracted from paraffin blocks and subjected to HPV 16 and 18 testing. The association between HPV 16 and 18 E6 oncoprotein and cell cycle proteins expression in ovarian carcinomas also was evaluated by immunohistochemistry.. We demonstrated that a majority of moderately and poorly differentiated ovarian carcinomas are characterized by strong expression of p53 and p16(INK4a) proteins. In contrast, strong staining with cyclin D1 antibody was observed in well differentiated tumors. The correlation between strong p53, pRb, Cyclin D1 and clinical stages of disease was also observed. We show that patients with high positivity for p53, p16(INK4a) and Cyclin D1 had a poor prognosis and reduced overall survival. The presence of HPV 16/18 DNA was detected in 17% of ovarian carcinomas. The tumor tissues that reacted positively to HPV E6 antibody in focal and diffuse manners had also significantly low p53 expression profile.. These findings suggest that p53, p16(INK4a) and Cyclin D1 expression and HPV infection may represent a promising tool toward the identification of ovarian cancer patients with poorer prognosis and shorter survival who might therefore need a more aggressive therapy and HPV screening.

Topics: Adolescent; Adult; Aged; Alphapapillomavirus; Antibodies, Monoclonal; Biomarkers, Tumor; Cell Cycle; Cell Cycle Proteins; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; DNA-Binding Proteins; Female; Human papillomavirus 16; Human papillomavirus 18; Humans; Immunohistochemistry; Middle Aged; Oncogene Proteins, Viral; Ovarian Neoplasms; Papillomavirus Infections; Prognosis; Repressor Proteins; Retinoblastoma Protein; Tumor Suppressor Protein p53

Human papillomavirus is involved in the carcinogenesis of tonsillar squamous cell carcinomas. Here, we investigated the expression and the prognostic value of key cell cycle proteins in the pRb and p53 pathways in both human papillomavirus type 16-positive and -negative tonsillar squamous cell carcinomas. Using immunohistochemistry, 77 tonsillar squamous cell carcinomas with known human papillomavirus type 16 status and clinical outcome were analyzed for expression of Ki67, p16(INK4A,) cyclin D1, pRb, p14(ARF), MDM2, p53, p21(Cip1/WAF1), and p27(KIP1). Results were correlated with each other and with clinical and demographic patient data. A total of 35% of tonsillar carcinomas harbored integrated human papillomavirus type 16 DNA and p16(INK4A) overexpression, both being considered essential features for human papillomavirus association. These tumors also showed the overexpression of p14(ARF) (P<0.0001) and p21(Cip1/WAF1) (P=0.001), and downregulation of pRb (P<0.0001) and cyclin D1 (P=0.027) compared with the human papillomavirus-negative cases. Univariate Cox regression analyses revealed a favorable survival rate for non-smokers (P=0.006), as well as for patients with T1-2 tumors (P<0.0001) or tumors showing low expression of cyclin D1 (P=0.028), presence of human papillomavirus and overexpression of p16(INK4A) (P=0.01), p14(ARF) (P=0.02) or p21(Cip1/WAF1) (P=0.004). In multivariate regression analyses, smoking and tumor size, as well as expression of cyclin D1 and p21(Cip1/WAF1), were found to be independent prognostic markers. We conclude that human papillomavirus positivity in tonsillar squamous cell carcinomas strongly correlates with p21(Cip1/WAF1) and p14(ARF) overexpression and downregulation of pRb and cyclin D1. In particular p21(Cip1/WAF1) overexpression is an excellent favorable prognosticator in tonsillar squamous cell carcinomas.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Cycle Proteins; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p21; Female; Gene Expression; Humans; Immunohistochemistry; Kaplan-Meier Estimate; Male; Middle Aged; Papillomaviridae; Papillomavirus Infections; Prognosis; Retinoblastoma Protein; Smoking; Tonsillar Neoplasms; Tumor Suppressor Protein p14ARF

High-risk human papillomavirus (hrHPV) infection is the major risk factor for cervical cancer (CxCa). The role of genetic susceptibility in the disease has been suggested, but the existing data lack consistency. We conducted a nested case-control study on 973 CxCa cases and 1,763 matched controls, from two Swedish population-based cohorts to examine the association of common genetic variants with CxCa risk. Human leukocyte antigen (HLA) alleles and 24 other polymorphisms in 14 genes were selected on the basis of reported association or mechanistic plausibility with an HPV infection or cervical cancer development. Genotyping was conducted using multiplex PCR and Luminex technology. A significant association of CxCa with various polymorphisms was observed: rs1800797 in the IL-6 gene (odds ratio [OR] = 0.88, 95% confidence intervals [CI]: 0.79-0.99); rs1041981 in the LTA gene (OR = 0.87, 95% CI: 0.78-0.98), and rs9344 in the CCND1 gene (OR = 1.14, 95% CI: 1.02-1.27), for those individuals carrying the rare allele. Additionally, the alleles 0401 and 1501 of the HLA class II DRB1 locus were associated with an increased risk (OR = 1.23, 95% CI: 1.04-1.45 and OR = 1.29, 95% CI: 1.11-1.50, respectively), and allele 1301 was associated with decreased risk (OR = 0.59, 95% CI: 0.47-0.73). The effects of CCND1 and the HLA*DRB1 alleles were independent of the effect of smoking. We did not find any association of risk with polymorphisms in genes related to the innate immune system. In conclusion, our study provides evidence for genetic susceptibility to CxCa due to variations in genes involved in the immune system and in cell cycle.

Topics: Adult; Aged; Case-Control Studies; Cervix Uteri; Cyclin D1; Female; Genotype; HLA-DR Antigens; HLA-DRB1 Chains; Humans; Interleukin-6; Middle Aged; Papillomaviridae; Papillomavirus Infections; Polymerase Chain Reaction; Polymorphism, Single Nucleotide; Risk Factors; Sweden; Uterine Cervical Neoplasms; Young Adult

In-depth study of cell cycle proteins and human papillomavirus (HPV) genotyping can provide useful information about the malignant potential of precursor lesions of cervical carcinoma (CC). Immunostaining of cell cycle-related proteins (p16, cyclin D1, Ki-67, p53, and ProEx C) was evaluated using tissue microarrays, and HPV genotypes were identified in 144 cervical tissue specimens encompassing normal or benign epithelial lesions, low- and high-grade squamous intraepithelial lesions (LSIL and HSIL, respectively), and CC. In addition, 14 cases with atypical immature metaplasia (AIM) were included to compare their immunohistochemical features with those of well-established precursor lesions. Expression of p16, Ki-67, and ProEx C was most associated with the severity of dysplasia. Positive expression of p16, Ki-67, and ProEx C and negative expression of p53 seem to be related to HPV-16 infection. AIM cases show an immunohistochemical pattern more similar to LSIL than to HSIL. Immunohistochemical assessment of cell cycle proteins may help to distinguish normal and benign conditions of the cervix from precursor lesions of CC.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Cell Cycle; Cell Cycle Proteins; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; DNA Topoisomerases, Type II; DNA-Binding Proteins; Female; Humans; Immunohistochemistry; Ki-67 Antigen; Minichromosome Maintenance Complex Component 2; Neoplasm Proteins; Nuclear Proteins; Papillomavirus Infections; Precancerous Conditions; Tissue Array Analysis; Tumor Suppressor Protein p53; Uterine Cervical Neoplasms

Human papilloma virus (HPV) was associated with sinonasal inverted papilloma (SIP) in 14/20 (70%) patients with a prevalence of HPV 6/11; alterations of the cell cycle proteins were statistically significant.. We investigated SIPs relationships between HPV infection and aberrant expression of cell cycle proteins.. Twenty SIPs were evaluated for p53, p16(INK4a), pRb, p21(WAF1), p27(Kip1), cyclin D1 and Ki-67 expression by immunohistochemistry. HPV was investigated by polymerase chain reaction (PCR).. HPV DNA was detected in 14/20 patients with inverted papillomas (IPs) (70%). The majority of tumours showed strong p16, p21, p27, pRb and cyclin D1 staining and little or no p53 expression. Tumours harbouring dysplasia were significantly more likely to be p53-positive and exhibit up-regulated p21 and p27, and showed altered intensity and distribution of reactive cells into and through the epithelium. Dysplastic epithelium was strongly reactive for p16 and the MIB 1 labelling index was almost 20%. These findings were associated with expression of p53 in the same zones. Comparing the p53 reactivity with the presence of HPV DNA, SIPs were stratified as follows: HPV + p53-, 12 (63.15%); HPV + p53+, 2 (10.52%); HPV - p53+, 3 (15.78%) and HPV - p53-, 2 (10.52%). Statistical analysis showed that HPV presence correlated with p53-positive immunostaining (p=0.045).

Topics: Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Cyclin-Dependent Kinase Inhibitor p21; Cyclin-Dependent Kinase Inhibitor p27; DNA Probes, HPV; Gene Expression Regulation; Gene Expression Regulation, Viral; Humans; Intracellular Signaling Peptides and Proteins; Ki-67 Antigen; Nose Neoplasms; Papilloma, Inverted; Papillomaviridae; Papillomavirus Infections; Paranasal Sinus Neoplasms; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Salivary Proline-Rich Proteins; Tumor Suppressor Protein p53

Estrogen stimulates human papilloma virus oncogene expression, promotes cervical cancer (CC) cell proliferation and prevents apoptosis. Therefore, blockage of estrogen function may have therapeutic application to CC.. CasKi CC cells were transfected with an adenovirus expressing a dominant negative estrogen receptor gene (Ad-ER-DN) and their responses were investigated by RT-PCR, Flow Cytometry and Western blot assays.. Transfected cells showed disturbance of cell colony morphology, reduced HPV E6 and E7 mRNA, interruption of cell proliferation, reduced cyclin D1 protein and expression of apoptosis.. We report, for the first time, the use of Ad-ER-DN to block estrogen receptors which led to dramatic changes in CC cells that are consistent with the possible reactivation of cellular p53 and Rb function. Their reactivation most likely allowed the recognition of existing chromosome abnormalities as a serious stress signal and the initiation of a cascade of cellular events in response to the stress, including the activation of the core apoptotic machinery which led to self-destruction of the CC cells.

Topics: Adenoviridae; Apoptosis; Cell Growth Processes; Cyclin D1; Female; Flow Cytometry; Genetic Therapy; Genetic Vectors; Human papillomavirus 16; Humans; Oncogene Proteins, Viral; Papillomavirus E7 Proteins; Papillomavirus Infections; Receptors, Estrogen; Repressor Proteins; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Transfection; Uterine Cervical Neoplasms

The histological criteria for cervical intraepithelial neoplastic lesions and their follow-ups have been established, but their reproducibility, specificity and sensibility are not certain. Immunohistochemical markers provide more information on each specific case, in order to facilitate its classification and, eventually, its prognosis. Using immunohistochemical techniques, this study analyzes the prognostic value of three markers (Ki-67, c-erbB2 and Cyclin D1) in cases of low grade squamous intraepithelial neoplasia (CIN-I), high grade squamous intraepithelial neoplasia (CIN-III), and infiltrating squamous cell carcinoma (SCC) taken from a group of cervical samples. In situ hybridization was performed in order to detect high-risk HPV. High risk HPV was demonstrated in 82%, 89% and 100% of the LGSIL, HGSIL and SCC cases, respectively. C-erbB2 expression was detected in 9%, 33% and 50% of the LSIL, HGSIL and SCC cases, respectively. The Ki-67 LI was 25%, 68% and 65.5% in the LGSIL, HGSIL and SCC cases, respectively. Nuclear Cyclin D1 expression was seen in 82%, 11% and 30% of the CIN-I,CIN-III and SCC cases, respectively. We observed that the cytoplasmic cyclin D1 expression increased with the severity of the lesion instead of the nuclear expression decreasing with the progression of the pathology. Nuclear and cytoplasmic Cyclin D1 expression seemed to be related to HPV high risk infection. We concluded that Cyclin D1, cerbB2 and The Ki-67 LI expression changed in relation to the severity of the lesion and that they could be helpful in making a differential diagnosis.

Topics: Antigens, CD1; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cyclin D1; Diagnosis, Differential; Female; Humans; Immunohistochemistry; In Situ Hybridization; Ki-67 Antigen; Papillomavirus Infections; Receptor, ErbB-2; Sensitivity and Specificity; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms

Inactivation of the retinoblastoma (pRB) pathway is a common event in oral squamous cell carcinoma particularly through the aberrant expression of the components within this pathway. This study examines the alterations of molecules within the pRB pathway by looking at the presence of homozygous deletions in p16(INK4A) and the expression patterns of pRB, cyclin D1 and CDK4, as well as the presence of human papillomavirus (HPV) in our samples. In our study, 5/20 samples demonstrated deletions of p16(INK4A) exon 1alpha. pRB overexpression was found in 20/20 samples, the expression was mainly observed in all layers of the epithelia, particularly in the basal layer where cells are actively dividing and aberrant pRB expression was found in 12/20 samples. Cyclin D1 and CDK4 overexpression was detected in 6/20 and 2/20 samples respectively in comparison to hyperplasias where both proteins were either not expressed or expressed at minimal levels (<10%). Strikingly, HPV was found to be present in all of our samples, suggesting that HPV plays a significant role in driving oral carcinogenesis. Notably, 17/20 of our samples showed more than one alteration in the pRB pathway, however, we did not find any significant relationship between the presence of HPV, homozygous deletion of p16(INK4A) and overexpression of pRB, cyclin D1 and CDK4. Collectively, this data demonstrates that alterations in the pRB pathway are a common event and involve the aberration of more than one molecule within the pathway. Furthermore, the involvement of HPV in all our samples suggests that HPV infection may play an important role in oral carcinogenesis.

Topics: Carcinoma, Squamous Cell; Cell Transformation, Neoplastic; Cyclin D1; Cyclin-Dependent Kinase 4; Cyclin-Dependent Kinase Inhibitor p16; Exons; Female; Gene Deletion; Homozygote; Humans; Immunohistochemistry; Malaysia; Male; Mouth Neoplasms; Papillomaviridae; Papillomavirus Infections; Retinoblastoma Protein; Tumor Suppressor Proteins

Small cell carcinomas (SmCCs) of the uterine cervix are rare tumors. The knowledge regarding protein expression of several checkpoint candidates of cell cycle regulation is limited. Surgically treated SmCCs were selected from our files for immunohistochemical staining (neuroendocrine markers, p53, p16, p14, and cyclin D1). Polymerase chain reaction analysis, using general primers, was performed for human papillomavirus analysis. Nine of 677 tumors (1.3%) were classified as SmCCs after Grimelius staining (8/9 tumors positive) and immunohistochemical reaction against neurone-specific enolase, chromogranin A, synaptophysin (7/9 positive tumors), and CD 56 (8/9 positive tumors). All specimens were positive for at least two of the above. Two SmCCs were p53 positive and one case was p14 positive. Cyclin D1 staining was completely negative. All cases showed strong nuclear and/or cytoplasmic p16-immunostaining. Seven tumors represented human papillomavirus positivity for high-risk types. Four patients died of the tumor after a median time of 36.7 months (range, 15-56 months), representing a 5-year survival rate of 56%. The results suggest that p16 is up-regulated or accumulated in the SmCCs of the uterine cervix, probably caused by infection with human papillomavirus. p14 inactivation is of high prevalence in SmCCs and detection rate of p53 is similar to other histologic types of cervical carcinomas.

Topics: Adult; Carcinoma, Small Cell; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Female; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Middle Aged; Papillomaviridae; Papillomavirus Infections; Polymerase Chain Reaction; Survival Rate; Tumor Suppressor Protein p14ARF; Tumor Suppressor Protein p53; Tumor Suppressor Proteins; Uterine Cervical Neoplasms

Pathogenetically, endometrioid adenocarcinomas of the endometrium are associated with hyperestrogenism and serous papillary carcinomas with alterations of p53. The etiology of primary endometrial squamous cell carcinoma (ESCC), however, is speculative. The purpose of this study was to evaluate the role of p14, p16, p53, cyclin D1, steroid hormone receptors, and human papillomaviruses (HPV) infection in the pathogenesis of primary endometrial squamous cell carcinoma. The expression of p16, p14, p53, cyclin D1, and steroid hormone receptors (estrogen, progesterone, and androgen) was examined immunohistochemically in 8 primary ESCCs. HPV analysis was performed using general primers and HPV typing. The median age of the patients was 62.1 years. Four cases showed positive nuclear and cytoplasmic p16 staining in an insular pattern, and 1 case nuclear positivity for p53 and estrogen receptors, respectively. Four of 8 cases were positive for progesterone receptor analysis and cyclin D1. All cases were negative for p14 and androgen receptor staining. All but one case were negative for HPV analysis. Five patients were alive with and without evidence of disease after a mean follow-up of 6.1 years. The results of this study suggest that alterations of the p16 pathway may play an etiologic role in at least a proportion of the ESCC, but without any association to HPV infection. Factors known to play a pathogenetic role in types 1 and 2 of endometrial carcinomas are not associated with primary ESCC. However, prognostically, ESCCs are more related to type 1 cancers.

Topics: Adult; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Squamous Cell; Cell Cycle Proteins; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Endometrial Neoplasms; Female; Humans; Middle Aged; Papillomaviridae; Papillomavirus Infections; Receptors, Steroid; Tumor Suppressor Protein p14ARF; Tumor Suppressor Protein p53

Human papillomavirus (HPV) plays a major role in the etiology of cervical cancer. However, a complex correlation between viral and cellular genes is necessary for cell cycle control deregulation in the progression to invasive cervical cancer (ICC). Cyclin D1 (CCND1) is an important positive regulator of the G1/S phase of the cell cycle. The CCND1 gene is located at 11q13 and is often altered in human cancers. We analyzed the A870G CCND1 polymorphism by polymerase chain reaction/restriction fragment length polymorphism analysis in 246 women including 50 cases with high-grade squamous intraepithelial lesions of the cervix (HSIL), 93 with ICC, and 103 healthy women. The GG genotype was associated with a 4.32-fold higher risk for the development of HSIL [adjusted odds ratio (aOR)=4.32, 95% confidence interval (CI) 1.50-12.46, P=0.0067), and a 3.26-fold increased risk for the development of ICC (aOR=3.26, 95% CI 1.42-7.53, P=0.006). The proportion of cervical cancer cases attributable to the GG CCND1 genotype was 17.26%. This study indicates that the A870G CCND1 polymorphism could act as a cofactor of HPV in the initiation of cervical carcinogenesis, particularly in the transformation zone of HPV-infected women, supporting evidence for a genetic factor in ICC risk.

Topics: Adult; Aged; Cyclin D1; Female; Genotype; Humans; Middle Aged; Papillomaviridae; Papillomavirus Infections; Polymorphism, Genetic; Risk; Uterine Cervical Neoplasms

Epidemiologic studies have implicated estrogenic exposure as well as human papilloma virus (HPV) infection in cervical carcinogenesis, and some studies have suggested that estrogen and HPV may play synergistic roles in cervical tumorigenesis. In this study, we report a novel finding that approximately 35% of cervical carcinomas tested (n = 19) express aromatase, the enzyme responsible for converting androgen to estrogen, the rate-limiting and final step in estrogen biosynthesis. On the other hand, no aromatase expression was detected in precancerous (n = 42) or normal cervical (n = 17) tissue samples. Increased aromatase was associated with increases in estrogen receptors (ER-alpha and ER-beta) and a decrease in progesterone receptor levels, suggesting that in situ estrogen signaling via ER may be involved in tumor growth. Stable overexpression of aromatase in HPV+ cervical cancer cells resulted in increased cellular proliferation, anchorage-independent growth, and ER expression and activity. In contrast, little change in ER was observed in HPV- cells. Steroid hormone receptor expression observed in vitro paralleled that seen in cervical carcinomas expressing aromatase. Aromatase overexpression also induced the expression of cyclin D1, proliferating cell nuclear antigen, and the HPV oncogenes, E6 and E7. Furthermore, the data underscores the importance of steroid receptor (estrogen and progesterone receptors) regulation in cervical carcinogenesis. To our knowledge, this is the first report demonstrating the induction of aromatase expression in cervical carcinomas, and opens the possibility that aromatase inhibitors may be potential therapeutic agents in cervical carcinomas expressing aromatase.

Topics: Aromatase; Cell Growth Processes; Cell Line, Tumor; Cervix Uteri; Cyclin D1; Enzyme Induction; Estrogen Receptor alpha; Estrogen Receptor beta; Estrogens; Female; Humans; Neoplastic Stem Cells; Papillomaviridae; Papillomavirus E7 Proteins; Papillomavirus Infections; Proliferating Cell Nuclear Antigen; Receptors, Progesterone; Uterine Cervical Neoplasms

infection by oncogenic subtypes of human papillomavirus (HPV) and cyclin D1 gene (CCND1) amplification are frequent events in head and neck squamous cell carcinomas. The objective of this paper is to establish the relationship between the presence of human papillomavirus (HPV) gene sequences and the development of CCND1 gene amplification in these tumours.. 59 squamous cell carcinomas of the head and neck were studied for HPV types 6b and 16 and CCND1 gene amplification by polymerase chain reaction.. HPV DNA was detected in 14 tumors (24%). Ten of them were positive for the HPV type 6b and 4 for the HPV type 16. CCND1 gene amplification was found in 15 cases (25%). Although we have found a higher frequency of CCND1 amplification in the HPV-positive cases (36%, versus 22% in the HPV-negative cases), these differences were not statistically significant (P= 0,32).. The presence of HPV gene sequences does not seem to be related to a significative higher incidence of CCND1 gene amplification in the squamous cell carcinomas of the head and neck.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cyclin D1; Gene Amplification; Head and Neck Neoplasms; Human papillomavirus 16; Human papillomavirus 6; Humans; Male; Middle Aged; Papillomavirus Infections

High-risk human papillomaviruses are the causative agents of cervical cancer and are also believed to be aetiologically involved in a subset of squamous cell carcinomas of the head and neck region, especially the tonsil. Cervical cancers arise through disruption of the pathways of p53 and the product of the retinoblastoma gene by the human papillomavirus oncoproteins E6 and E7. It is generally assumed that the same pathways are involved in human papillomavirus-induced carcinogenesis at other mucosal surfaces. However, the patterns of expression of cell cycle proteins targeted by human papillomavirus E6 and E7 in cancers from different anatomic sites have been inconsistent, due to either biologic or technological factors. In this study, 73 human papillomavirus, 16-positive cervical squamous cell carcinomas (35 from Australian and 38 from Chinese women) were analysed for the expression of p53, pRb, p16(INK4A), p21(CIP1/WAF1), p27(KIP1) and cyclin D1 by semiquantitative immunohistochemistry. Cervical cancers from Chinese women were found to be significantly more likely to overexpress p53, pRb, p21 and p27 than their Australian counterparts. These findings were compared with those from 31 human papillomavirus 16-positive tonsillar squamous cell carcinomas, all of Australian origin, tested using the same methodology. Comparisons of the tonsillar and combined cervical data showed that tonsillar cancers were significantly more likely to be p53-positive, whereas cervical cancers were significantly more likely to overexpress pRb, p16 and p27. When the tonsillar data were compared with cervical data from Australian women, the associations for p53 and pRb remained. These findings represent new evidence that the molecular pathways to human papillomavirus-induced mucosal cancer may be influenced by anatomic location and ethnicity.

Topics: Adult; Aged; Aged, 80 and over; Australia; Carcinoma, Squamous Cell; Cell Cycle Proteins; China; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p16; Cyclin-Dependent Kinase Inhibitor p21; Cyclin-Dependent Kinase Inhibitor p27; Female; Humans; Immunohistochemistry; Middle Aged; Neoplasm Staging; Papillomaviridae; Papillomavirus Infections; Retinoblastoma Protein; Tonsillar Neoplasms; Tumor Suppressor Protein p53; Tumor Suppressor Proteins; Uterine Cervical Neoplasms

Many investigators have studied the expression of G1 phase regulatory protein in uterine cervical cancer. However, it is unclear which step of the genetic expression participates in cyclin D1 expression and what its prognostic meaning is. The aims of this study were to evaluate the regulatory level of cyclin D1 expression and the relationship between the expression of cyclin D1 and its inhibitor, p21WAF1/CIP1, and to evaluate their impact on the prognosis of early stage cervical cancer.. The presence of cyclin D1 mRNA was studied using Northern blot in 6 normal cervices and 7 invasive cervical cancer specimens. Western blot was used to detect the cyclin D1 protein in 8 normal cervices and 8 invasive cancer specimens. Thirty-two cases of FIGO stage Ib-IIa cervical cancers (28 squamous cell carcinomas, 3 adenocarcinomas, 1 adenosquamous cell carcinoma), 31 cases of cervical intraepithelial neoplasia 3 (CIN 3), and 28 normal cervices were stained for cyclin D1 and p21(WAF1/CIP1) using monoclonal antibody. Statistical analysis was performed to assess the differences in expression and their prognostic significance. RESULTS. Cyclin D1 mRNA was found to be underexpressed in cervical cancer. Western blot also revealed underexpression of cyclin D1 protein in cervical cancer compared to normal controls. Positive immunohistochemical staining of cyclin D1 was noted in 28/28 (100%) of the normal controls, 1/31 (3%) cases of CIN 3, and 9/32 (28%) cases of invasive cancer. The number of positively stained specimens was lower than that of normal controls in CIN 3 and cervical cancer specimens (P = 0.005). Fifteen of 28 (54%) normal controls, 15/31 cases (48%) of CIN 3, and 27/32 cases (84%) of invasive cancer were proved positive for p21WAF1/CIP1 immunohistochemistry. p21WAF1/CIP1 was more highly expressed in cervical cancer than in that of either normal controls or CIN specimens (P = 0.001). Positive immunostaining of cyclin D1 and p21WAF1/CIP1 was not related to high-risk factors (pelvic lymph node metastasis, deep cervical stromal invasion, parametrial invasion, large tumor size, and unusual histologic type) and human papilloma virus infection. Positive cyclin D1 immunostaining was associated with decreased disease-free survival and a lower overall survival (P = 0.0175 and 0.0189, respectively). On multivariate analysis, positive cyclin D1 expression was a significant prognostic variable for recurrence (P = 0.0004).. Underexpression of cyclin D1 was regulated at the level of transcription in cervical cancer. Although cyclin D1 was underexpressed in cervical neoplasias, it was more frequently expressed in malignant lesions. p21WAF1/CIP1 was more highly expressed in cervical cancers than in either normal cervices or CIN 3 specimens. Unfavorable prognoses were associated with cyclin D1 expression, and not with the expression of its inhibitor, p21WAF1/CIP1.We conclude that immunohistochemical assessment of cyclin D1 can be a useful molecular marker for predicting prognosis in early stage cervical cancer of the uterus.

Topics: Blotting, Northern; Blotting, Western; Cyclin D1; Cyclin-Dependent Kinase Inhibitor p21; Cyclins; Female; Gene Expression; Humans; Immunohistochemistry; Neoplasm Recurrence, Local; Neoplasm Staging; Papillomaviridae; Papillomavirus Infections; Prognosis; RNA, Messenger; Survival Analysis; Tumor Virus Infections; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms

Human papillomavirus infection has been suggested to play a role in the development of epithelial carcinomas, particularly those of the uterine cervix. Less information is available on the role of the virus in oral lesions. It has been proposed that the viral oncoproteins specifically complex with the products of cellular tumor suppressor gene, namely E6 with p53 and E7 with retinoblastoma gene product. Inactivation or mutation in p53 gene is also known to result in loss of control over the cell cycle and increases in tumor proliferation rates. The present study examines the role of HPV infection in relation to p53 and the extent of the tumor proliferative compartment reflected by cyclin D1 and Ki-67 expression during various phases of tumor progression in the oral epithelium.. Nonisotopic in situ hybridization (NISH) was performed to detect HPV 6/11 and 16/18. Expression of p53, cyclin D1, Ki-67, and the HPV 16/18 E6 protein were detected by immunocytochemistry.. There was significant correlation between the extent of histological abnormality and HPV infection. A correlation (r = 0.250, P = 0.0089) was evident between the presence of HPV 16 and occurrence of invasive cancer. Expression of the tumor suppressor p53 protein also showed significant positive correlation with histology (r = 0.475, P = 0.00004). The tumor proliferative fraction also increased with the extent of histological abnormality (r = 0.387, P = 0.0003 for cyclin D1 and r = 0.463, P = 0.0001 for Ki 67). Accumulation of p53 and increase in tumor proliferation also correlated to the presence of HPV infection (r = 0.511, P = 0.00003 for p53; r = 0.478, P = 0.00002 for cyclin D1 and r = 0.521, P = 0.00004 for Ki-67).. The present study thus demonstrates the importance of HPV infection in oral tissue. Expression of the high-risk HPV 16/18 E6 protein also appears to be a critical event along with aberrant p53 expression. These results are of significance to the molecular epidemiology of oral cancer and may also be used to supplement and elaborate the diagnosis of oral lesions.

Topics: Cyclin D1; Genes, p53; Humans; Immunohistochemistry; In Situ Hybridization; Ki-67 Antigen; Mouth Mucosa; Mouth Neoplasms; Oncogene Proteins, Viral; Papillomaviridae; Papillomavirus Infections; Tumor Virus Infections

Both SV40 and JC virus (JCV) appropriate the host cell replicative machinery to attend to their own reproductive needs. SV40 large T antigen is able to induce the expression of cyclins A, B1, and E (but not of cylin D1) in transfected diploid cells. Whether JCV infection influences cyclin expression in a similar fashion in the setting of progressive multifocal leukoencephalopathy (PML) remains unknown. Brain lesions from 7 PML cases (4 autopsies and 3 biopsies) were immunohistochemically investigated for the expression of Ki-67 and cyclins A, B1, and D1. All 7 cases showed strong positivity for Ki-67 and cyclins A and B1 in JCV-infected oligodendrocytes and astrocytes, the nuclear immunolocalization of cyclin A being in strong contrast to the cytoplasmic distribution of cyclin B1. No immunostaining for cyclin D1 was obtained in any of the 7 cases. These findings suggest that JCV infection is associated with overexpression of Ki-67 and cyclins A and B1 in PML host glial cells. Since cyclin changes in JCV-infected cells recapitulate SV40 T antigen-associated cyclin fluctuations, it appears reasonable to think that JCV T antigen shares some of the previously described capabilities of SV40 T antigen to alter cyclin expression for the sake of viral replication.

Topics: Acquired Immunodeficiency Syndrome; Adult; Brain; Cyclin A; Cyclin B; Cyclin B1; Cyclin D1; Cyclins; DNA, Viral; Humans; Immunocompromised Host; Immunoenzyme Techniques; In Situ Hybridization; JC Virus; Ki-67 Antigen; Leukoencephalopathy, Progressive Multifocal; Male; Middle Aged; Papillomavirus Infections; Tumor Virus Infections

In this study, we have demonstrated the overexpression of cyclin D1 protein in 24 (92%) of 26 low-grade squamous intraepithelial lesions infected with low-risk human papillomaviruses (HPV 6, 11, 42, 43, and 44) and the absence of cyclin D1 expression in 25 (87%) of 29 lesions infected with high-risk HPVs (HPV 16, 18, 31, 33, 39, 45, 51, 52, 56, 58, and 66). The expression of cyclins E, A, and B proteins was increased in both low-risk and high-risk HPV infections. Tetrasomy of chromosomes 1, 3, 11, 17, 18, and X was present in nine lesions, all of which were infected with high-risk HPVs, but was not related to the pattern of cyclin expression. These data provide in vivo evidence that low- and high-risk HPV types alter cell cycle control by different mechanisms and that cell cycle checkpoint abnormalities are induced by high-risk, but not low-risk, HPV infection.

Topics: Cell Cycle; Chromosome Aberrations; Chromosome Disorders; Cyclin D1; Epithelium; Female; Humans; Papillomaviridae; Papillomavirus Infections; Tumor Virus Infections; Uterine Cervical Dysplasia; Uterine Cervical Neoplasms; Warts

Head and neck squamous cell carcinomas (SCCs) seem to follow a multistep process of carcinogenesis in which chemical and/or viral agents are associated with specific genetic alterations. The prevalence of human papillomavirus (HPV) infection and the amplification of the cyclin D1 (CCND1) gene were evaluated in a series of 75 laryngeal SCCs by PCR with HPV consensus primers and Southern blot analysis with a CCND1-specific probe, respectively. HPV DNA was detected in 22 of 75 (29.3%) tumors, and it belonged almost exclusively to the highly oncogenic HPV-16, HPV-18, and HPV-33. CCND1 gene amplification was found in 15 of 75 (20%) tumors, and it was associated with HPV infection in a statistically significant manner (chi2 = 20.3; P < 0.001). Because the viral oncoproteins E6 and E7 from high-risk HPV types are known to promote genomic rearrangements, these findings suggest that amplification of the CCND1 gene in laryngeal SCCs may occur as a consequence of the genomic instability associated with HPV infection. In turn, amplified CCND1, either alone or in conjunction with a direct action of the viral oncoproteins E6 and E7, could lead to a perturbation of the cell cycle. This model could explain the involvement of high-risk HPV types in laryngeal carcinogenesis.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Squamous Cell; Cyclin D1; DNA, Viral; Female; Gene Amplification; Humans; Laryngeal Neoplasms; Male; Middle Aged; Papillomaviridae; Papillomavirus Infections; Tumor Virus Infections