Folate and choline metabolism gene variants in relation to ovarian cancer risk in the Polish population.

Data indicates that genetic factors alone do not account for ovarian tumorigenesis, suggesting that epigenetic status additionally affects this process. Therefore, we assessed the possible contribution of polymorphic variants of genes that may affect DNA methylation to the risk of ovarian cancer incidence in the Polish population. Using PCR-RFLP and HRM analyses, we studied the distribution of BHMT (rs3733890), MTHFD1 (rs2236225), MTHFR (rs1801133), MTR (rs1805087), MTRR (rs1801394) and TCN2 (rs1801198) genotypes and alleles in patients with ovarian cancer (n = 136) and controls (n = 160). Moreover, using DNA and methylation-specific PCR (MSP) we also determined the methylation of the Cadherin 13 (CDH13) promoter in cancerous tissue from these patients. We did not observe a significant association between all studied gene variants and the incidence of ovarian cancer. The lowest P (trend) = 0.1226 was observed for the MTHFR Ala222Val polymorphism. Moreover, the lowest P = 0.0772 was found in the comparison of MTHFR Ala/Ala versus Val/Val and Val/Ala genotypes in patients and control groups. The multifactor dimensionality reduction analysis also did not indicate a significant interactive genetic effect on ovarian cancer incidence for all analyzed SNPs. However, we observed frequent methylation of the CDH13 promoter in approximately 21% (29/136) patients with ovarian carcinomas. Our results might suggest that the selected polymorphic gene variants may not contribute to ovarian cancer incidence.

[Preinvasive vulvar and cervical cancer in a 32-year-old woman, DNA HPV 16 positive with mtDNA mutation--case study]. / Rak przedinwazyjny sromu i szyjki macicy u 32-letniej kobiety DNA HPV 16 pozytywnej z mutacja mtDNA--opis przypadku.

Coincidence of preinvasive vulvar and cervical cancer in young women is very rare. Lesions like VIN 3/preinvasive vulvar cancer and CIN 3/preinvasive cervical cancer are strictly connected with viral infection and are multilocular. In the presented case the following tests have been performed: HPV DNA test for the presence of 13 oncogenic HPV types, mRNA HPV test for the presence of transcripts for HPV 16, 18, 31, 33, 45 and the analysis of mtDNA D-Loop region. In the examined patient HPV 16 infection, as well as the presence of transcripts for HPV 16 E6/7 were diagnosed. The analysis of mtDNA D-Loop region showed nucleotide lesions like: T>C 16.192, T>C 16.223, T>C 16.292, C>T 16.325, C>T 16.579.

[Assesment of real optoelectronic method in the detection of cervical intraepithelial neoplasia]. / Ocena przydatnosci metody optoelektronicznej w wykrywaniu sródnablonkowej neoplazji szyjki macicy.

INTRODUCTION: Biophysics is becoming an important factor in prevention of cervical cancer. One of the most promising concepts of biophysical program for the prevention of cervical cancer is the optoelectronic method--Truscreen. OBJECTIVE: The objective of the work was to determine the usefulness of the real optoelectronic device in the detection of cervical intraepithelial neoplasia MATERIAL AND METHOD: From August 2006 till September 2007, 147 patients were examined in the Laboratory of Pathophysiology of Uterine Cervix in Gynecology and Obstetrics Clinical Hospital of Karol Marcinkowski University of Medical Sciences. The study reports prospective, blind trial. The following diagnostic examinations were carried out: examination with the use of optoelectronic method, DNA HPV HIGH RISK test, colposcopy examination and histopatologic biopsy. RESULTS: The specificity of the optoelectronic method for positive diagnoses amounted to 84%. The sensitivity for intraephitelial changes (CIN 1, CIN 2) was estimated at 53%, and for severe grades (CIN 3, Carcinoma planoepitheliale) at 80%. CONCLUSIONS: Optoelectronic diagnostics is useful in the detection of cervical neoplastic changes. The optoelectronic method may prove to be a significant diagnostic device at the basic and advanced level of cervical screening. The advantage of the optoelectronic method over prevention based on cytodiagnostics and colposcopy, includes the immediate result of the examination and its automation as well as standardization.

Analysis of hMLH1 and hMSH2 expression in cisplatin-treated ovarian cancer patients.

BACKGROUND: Loss of DNA mismatch repair may result in resistance to platinum- based anticancer drugs. The hMLH1 and hMSH2 proteins play a critical role in the maintenance of genome integrity and are involved in resistance to platinum-based therapy in colorectal cancer, which is deficient in hMLH1 protein and endometrial cancer, as well as in hMSH2 protein. However, the predictive value of MLH1 and MSH2 expression in ovarian cancer cisplatin-resistance is still to be determined. OBJECTIVE: The aim of this study was to investigate the expression of hMLH1 and hMSH2 proteins in ovarian carcinoma specimens and to evaluate their prognostic significance by means of overall survival (OS) and progression-free survival rates (PSF). MATERIAL: Ovarian cancer tissues were obtained from 61 patients: 45 platinum-sensitive and 16 platinum-resistant. hMLH1 and hMSH2 proteins expression was evaluated by immunohistochemistry, with the use of mouse monoclonal antibodies clone 14 for hMLH1 and clone FE11 for hMSH2. The log-rank test and Kaplan-Meier statistics were used to analyze the relationship between proteins expression and progression free survival, as well as the overall survival. RESULT: No significant correlation was found between hMLH1 and hMSH2 expression and overall survival and progression free survival in the group of patients sensitive and resistant to cisplatin. No significant difference was found in proteins expression intensity between the two compared groups of patients. Age of patients, type of cancer histology, FIGO staging, grading, clinical response and CA 125 did not reveal correlation with the expression of the analyzed proteins. CONCLUSION: The immunohistochemical expression of hMLH1 and hMSH2 proteins in ovarian cancer has no predictive value in resistance to cisplatin.

The 3020insC NOD2 gene mutation in patients with ovarian cancer.

OBJECTIVE: There is an increasing evidence that genetic factors play a role in the etiology of malignant tumors. Mutations of BRCA1 and BRCA2 genes are responsible for an increased risk of ovarian cancer. The role of mutations in NOD2 gene in this type of neoplasm is still under investigation. THE AIM: The aim of this study was to determine: 1. incidence of NOD 2 3020insC constitutional mutation in a group of consecutive women with ovarian cancer, 2. risk of developing ovarian cancer in patients with NOD2 gene mutation, 3. clinical and pathological features of ovarian cancer in NOD2 gene mutation carriers. PATIENTS AND METHODS: Clinical and pathological data were collected from 257 non-selected patients with primary epithelial ovarian cancer. The researches identified NOD2 3020insC gene mutation. On the basis of patient source documentation we obtained the data concerning the age of patients at diagnosis, histopathological recognition, FIGO stage and morphological grade G. RESULTS: 19 out of 257 women were identified with germ-line 3020insC mutation of NOD2 gene (7.39%). An increased risk of ovarian cancer in NOD2 mutation carriers was not revealed (OR=1.01; p=0.928; 95% Cl=0.61-1.66). The mean age at diagnosis of patients with NOD2 mutation was 54.8 (SD=9.9), while for non-carriers it was 53.2 (SD=10.2). The difference between these frequencies was statistically irrelevant (p=0.550). Clinical and pathological profile of ovarian cancer was made. We assessed the following features: age at disease onset, histopathology, FIGO stage and morphological grade G. For NOD2 mutation carriers no statistically significant features of ovarian cancer were revealed. CONCLUSION: 1. Despite high frequency of constitutional mutations occurrence in NOD2 gene in women with ovarian cancer, genetic testing seem not to be justified in all women diagnosed with this disease. 2. Due to a lack of increased risk of ovarian cancer in NOD2 gene mutation carriers, proceedings for them may not differ from recommendations for general population. 3. It is difficult to determine characteristic clinical and pathological features of ovarian cancer for NOD2 gene mutation carriers.

[Agresive variants of uterine cervical cancers]. / Agresywne warianty raka szyjki macicy.

Three different variants of biologically agressive cervical cancers, such as: basaloid carcinoma, glassy cell carcinoma and small cell endocrine carcinoma, have been reported. Hematoxylin and eosin, periodic acid-Schiff and immunohistochemical stains were used. The course of disease and aplication of treatment has been analyzed as well. Characteristic morphologic features of these cancers have been emphasized. Rather limited susceptibility for radiotherapy and chemiotherapy of these cancers has not gone unnoticed. The description of these types of cervical cancers might enable us to arrive at right conclusions, both clinical and diagnostic.

[Diagnosis of papilloma viruses in cervical cancer in 414 women from Wielkopolska region by the immunohistochemical assessment]. / Identyfikacja wirusów brodawczaka w 414 przypadkach raka inwazyjnego szyjki macicy u kobiet regionu wielkopolskiego w powiazaniu z badaniami immunohistochemicznymi.

OBJECTIVES: Cervical cancer in Poland is second most common type of cancer, after breast cancer. There are known risk factors of cervical cancer development and the most serious one is human papilloma virus infection (HPV). DESIGN: The aim of our paper is present the result study 414 cervical cancer of women from Wielkopolska region treated at the Department of Gynecology and Obstetrics, Karol Marcinkowski University of Medical Sciences in Poznan. MATERIALS AND METHODS: In morphological study grading, staging and vascular invasion were estimated. In cervical cancer tissue papillomaviruses PCR method were used. In immunohistochemical study, expression of proteins checking the cell cycle, share in signal transduction to nucleus, cell receptors for steroid hormones and viruses oncogenic proteins were investigated. In the part of cancer gene mutation of p53 (60 cancers) i k-RAS (40 cancers) were searched. RESULTS: In cancers HPV 16/18 infected vascular invasion were more frequently (p < 0.013). No statistically significant difference in cellular proteins expression in the HPV16/18 positive cancers, HPV16/18 negative and cancers without HPV was observed. However significant difference were demonstrated in proteins expression depending from degree of cancer stage. CONCLUSIONS: The result of these studies suggest that super expression for EGFr is poor prognostic factor in the early stage of cancers (I-II0).

Expression of neuroendocrine markers in endometrial carcinomas - an immunohistochemical analysis.

The main aim of this study was the analysis of undifferentiated cancers and mixed ones positive for neuroendocrine biomarkers without morphological features - typical for this type of cancer. The obtained results indicated the necessity for neuroendocrine marker tests in undifferentiated cancers and mixed ones. Immunopositive results revealed enhanced malignancy of these cancers and the necessity for additional chemotherapy.

Transcriptional analysis of CXCR4, DNMT3A, DNMT3B and DNMT1 gene expression in primary advanced uterine cervical carcinoma.

The development of cervical cancer requires genetic and epigenetic factors which result in the persistence of a malignant phenotype. Cervical cancer exhibits also some unique differences from other solid tumors. Normal cervical stratified epithelia have characteristics of hypoxic tissue with over-expression of HIF-1 (hypoxia-inducible factor-1) transcription factor, which targets the transcription of over 70 genes involved in many aspects of cancer biology. One of the genes, which could be induced by HIF-1 is chemokine (C-X-C motif) receptor 4 (CXCR4). CXCR4 could also be epigenetically regulated by methylation of CpG dinucleotides located in the promoter region. Here, we examined the CXCR4, DNMT3A, DNMT3B and DNMT1 transcript levels in cancer tissue (n=30) and non-cancer, normal uterine cervical tissue (n=30) from a Polish cohort. We also compared the methylation status of CXCR4 promoter region in cancer and normal tissue samples. Our result showed significantly higher levels of CXCR4, DNMT3A, DNMT3B and DNMT1 transcript (p=0.0058, 0.0163, 0.0003 and <0.0001, respectively) levels in cancer tissue as compared to normal samples. We did not observe DNA methylation in the CXCR4 promoter region in either control or cancer tissue samples. CXCR4 has a functional hypoxia response element (HRE) in the promoter region, located -1.3 kb from the transcription start site. Our work shows for the first time that HIF-1A could promote the induction of CXCR4 gene expression (Spearman's correlation coefficient = 0.515, p=0.003) in patients with primary advanced uterine cervical carcinoma.

Carcinosarcoma (malignant mixed mesodermal tumor) of the uterus: clinicoimmunohistochemical and histogenetic characteristics.

To search for favourable prognostic factors in carcinosarcoma (CS) on the basis of clinical, morphological and immunocytochemical data, while simultaneously considering the histogenesis of this neoplasm. Thirty two uterine CS patients were analysed based on clinical and morphological data. In addition, each specimen was examined by immunohistochemistry with antibodies characteristic for relevant types of cells and tissues. The presence of both carcinomatous and sarcomatous patterns was observed in all tumours. Among carcinomatous patterns, endometrioid carcinoma was the commonest, while serous, clear cell, and undifferentiated carcinomas were less common. Among sarcomatous patterns, endometrioid sarcomas represented the largest group, while leiomyosarcomas, chondrosarcomas, fibrosarcomas, osteosarcomas, and rhabdosarcomas were rarely observed. Mitotic activity was evidently higher in carcinomas. In seven cases, the expression of both cytokeratin and vimentin was noted in cells of carcinomatous patterns. We found that an early diagnosis (stage I-II) and an initially aggressive surgical cytoreduction were favourable prognostic factors in CS. Furthermore, the presence of cytokeratin-vimentin positive cells in carcinomatous patterns suggests sarcomatous metaplasia of adenocarcinoma. However, the prognostic value of various histological structures of carcinosarcomas could not be identified.

Increased expression of HIF-1A and its implication in the hypoxia pathway in primary advanced uterine cervical carcinoma.

The development of cervical cancer exhibits some unique differences compared to other solid tumors. Normal cervical stratified epithelia have characteristics of hypoxic tissue. Lack of oxygen (hypoxia) induces the HIF-1 (hypoxia-inducible factor-1) transcription factor, which is a heterodimer composed of a constitutively expressed ß subunit and a hypoxia-inducible α-subunit. HIF-1A targets the transcription of over 70 genes involved in many aspects of cancer biology. In well-oxygenated environments, the HIF-1A subunit is hydroxylated, recognized and marked for proteosomal destruction by an E3 ubiquitin ligase, the von Hippel-Lindau protein (pVHL) complex. Under hypoxic stress, proline hydroxylase (PHD) activity is diminished, and stabilized HIF-1A is involved in the activation of the tissue response to hypoxia. Here, we examined the HIF-1A and VHL transcript levels and HIF-1A protein levels in cancerous tissue (n=30) and non-cancerous, normal uterine cervical tissue (n=30). We also compared the methylation status of HIF-1A and of the VHL promoter regions in cancerous and normal tissue samples. Significantly higher levels of HIF-1A and VHL transcripts (p<0.0001 and p=0.0042, respectively) and of HIF-1A protein (p=0.0037) were detected in cancerous tissue compared to normal samples. We did not observe DNA methylation in the HIF-1A and VHL promoter region in either control or cancerous tissue samples. VHL has a functional hypoxia response element (HRE) in the promoter region, and the induction of this HRE acts within a negative feedback loop to limit the hypoxic HIF-1A response. Our findings may suggest that HIF-1A could promote its own degradation by the induction of VHL gene expression (Spearman correlation coefficient, 0.515; p=0.003). Our study shows for the first time that this increase in VHL expression could be HIF-1A-dependent and serves within a negative feedback pathway during hypoxia to regulate the cell-specific oxygen threshold for HIF-1A activation.

Reduced expression of PHD2 prolyl hydroxylase gene in primary advanced uterine cervical carcinoma.

Decreased PHD2 expression in human carcinomas has been considered a critical factor in supporting tumor angiogenesis and growth. We studied the levels of PHD2 transcript and protein in advanced cervical cancer specimens (n=27) and normal uterine cervical tissue samples (n=27). Real-time quantitative PCR and Western blotting analysis showed significantly lower levels of PHD2 transcript (P=0.0088) and protein (P=0.0095) in cancerous tissues as compared to corresponding normal tissue. Using DNA sequencing analysis, we also found an accumulation of mutations in promoter regions of PHD2 in advanced cervical cancer specimens. Moreover, computer analysis of these mutations showed a loss of binding sites for many transcription factors. Our results suggest PHD2 as a possible target in anti-angiogenic therapies in advanced uterine cervical carcinoma.

Serum and tissue levels of insulin-like growth factor-I in women with dysplasia and HPV-positive cervical cancer.

The ectocervical epithelium is the target of the oncogenic human papillomavirus (HPV), which acts as an etiological agent in the development of cervical carcinoma. However, the HPV-mediated transformation of human epithelial cells is a multi-step process dependant on unknown factors additional to the virus, which is a necessary but, in and of itself, insufficient catalyst. In the present study, we characterized the role of insulin-like growth factor-I (IGF-I) in this process. IGF-I is an endocrine hormone with an autocrine and paracrine role in many tissues. Our data demonstrated that autocrine secretion of IGF-I can contribute to HPV-induced carcinogenesis of the epithelium. An immunohistochemical study showed that IGF-I was present in the nuclei of the reproductive layer of the paraepidermal epithelium in 89% of cases of intraepithelial neoplasia (CIN I-III) and 60% of cases of invasive cervical cancer. The presence of IGF-I at a nuclear localization in the cells studied suggests that it may also have intranuclear actions.