Polymorphic variants in vitamin D signaling pathway genes and the risk of endometriosis-associated infertility.

It has recently been reported that vitamin D blood plasma levels are associated with reduced risk of endometriosis. The present study aimed to investigate whether the vitamin D binding protein (GC), vitamin D receptor (VDR), and retinoid X receptor (RXR) gene variants may be genetic risk factors for endometriosis­associated infertility. The subjects consisted of 154 women with endometriosis­associated infertility and 347 controls. Using polymerase chain reaction restriction fragment length polymorphism and high resolution melt techniques, the GC rs1155563, rs2298849 and rs7041; RXRA rs10881578, rs10776909 and rs749759; VDR BsmI rs1544410; and FokI rs2228570 single nucleotide polymorphisms (SNPs) were investigated in the patients with endometriosis and the healthy controls. The results indicated that no significant differences were observed between the genotype and allele frequencies of all experimental SNPs in the vitamin D signaling pathway genes in women with endometriosis-associated infertility and controls. However, a significant association was present between the A­T haplotype, consisting of VDR rs1544410 and rs222857 minor alleles, and endometriosis-associated infertility [OR=1.659 (1.122­2.453), P=0.011]. The results of the present study suggested that VDR gene variants act as genetic risk factors for endometriosis­associated infertility.

EVC gene polymorphisms and risks of isolated hypospadias - a preliminary study.

INTRODUCTION: Hypospadias has a complex etiology with both genetic and environmental factors contributing to the condition. Urogenital abnormalities including hypospadias, are found in 22% of cases with Ellis van Creveld syndrome (EvC). Mutations in the EVC gene can cause major and minor anomalies, which form phenotypes that partially overlap with those present in EvC. The aim of this study was to evaluate the association between nucleotide variants of the EVC gene and the risk of hypospadias. MATERIAL AND METHODS: Four single nucleotide polymorphisms (SNPs) of the EVC gene (rs3774856, rs2302075, rs1383180, rs7680768) were taken under investigation in 96 patients with isolated hypospadias and 284 matched controls. Genotyping of all polymorphisms was carried out by PCR and followed by appropriate restriction enzyme digestion (PCR-RFLP). RESULTS: Individuals homozygous for the SNP rs2302075 (p.Thr449Lys) showed an elevated risk for hypospadias. Haplotypes containing the rs2302075 variant also revealed modest associations with hypospadias, which did not survive multiple testing corrections. None of the other tested EVC polymorphisms displayed significant association with the risk of hypospadias, either in dominant or recessive inheritance models. CONCLUSIONS: The results of this study suggest that polymorphic variants of the EVC gene do not substantially contribute to the risk of hypospadias based on our study population. However, further studies should help to clarify the relationship between polymorphisms of EVC and hypospadias.

Murine Double-Minute 2 Homolog Single Nucleotide Polymorphisms 285 and 309 in Cervical Carcinogenesis.

BACKGROUND AND OBJECTIVE: In Caucasians, the MDM2 single nucleotide polymorphism (SNP) 285 G>C (rs117039649) neutralizes the effect of 309 T>G (rs2279744), which increases MDM2 expression and impairs the p53 pathway. In this study, we examined the distribution of these two SNPs in Polish women with squamous cell carcinoma (SCC) (n = 379), adenocarcinoma (n = 59) and other cervical tumor types (n = 18). METHODS: The polymerase chain reaction-restriction fragment length polymorphism technique and DNA sequencing were employed in our study. RESULTS: The P trend value calculated for the MDM2 285 G>C polymorphism was statistically significant (P trend = 0.016) for SCC. Using logistical regression analysis adjusted for the effect of age, pregnancy, oral contraceptive use, tobacco smoking, and menopausal status, we observed that the MDM2 285 G>C SNP protected against SCC, with an adjusted odd ratio (OR) for the C carriers versus G/G genotype of 0.536 (P = 0.019). Stratified analyses of MDM2 285 G>C revealed a protective role of the C allele against SCC in women with a positive history of oral contraceptive use (age-adjusted OR 0.413, P = 0.021) and in premenopausal women (age-adjusted OR 0.362, P = 0.022). We also found that the 285GG/309GG vs 285GG/309 TT genotype increased the risk of SCC (adjusted OR 1.890, P = 0.005). However, the 285CC/309GG + 285GC/309GG versus 285GG/309GG genotype reduced the risk of SCC (adjusted OR 0.311, P = 0.004). CONCLUSION: Our results demonstrate that the MDM2 285C gene variant and 285CC/309GG + 285GC/309GG genotypes protect against SCC, most likely by neutralizing the effect of the 309 T>G SNP. The 285GG/309GG genotype increases the risk of SCC possibly due to increased MDM2 expression.

Polymorphic variants in the dopamine receptor D2 in women with endometriosis-related infertility.

Data suggests that dopamine receptor DRD2 gene variants may contribute to hyperprolactinemia and that they may be risk factors for endometriosis-related infertility. The purpose of the present study was to determine whether nucleotide variants of the DRD2 gene may be associated with infertility related to endometriosis. Five DRD2 SNPs, rs1800497, rs6277, rs2283265, rs4245146 and rs4648317, which are located in different blocks of linkage disequilibrium, were studied in 151 cases and 381 controls. No significant differences between DRD2 rs1800497, rs6277, rs2283265, rs4245146 and rs4648317 genotype, allele nor haplotype frequencies were observed in women with endometriosis-related infertility compared with the control group. The present results did not confirm DRD2 gene variants to be genetic risk factors for endometriosis-related infertility.

Stromal cell-derived factor-1 G801A polymorphism and the risk factors for cervical cancer.

Although certain studies have demonstrated no association between the stromal cell­derived factor­1 (SDF1­3') G801A single nucleotide polymorphism (SNP) and cervical carcinoma, the interactions between the SDF1­3' G801A SNP and contraceptive use, menopausal status, parity and tobacco smoking remain to be fully elucidated. Using polymerase chain reaction­restriction fragment length polymorphism, the distribution of SDF1­3' G801A genotypes in patients with cervical cancer (n=462) against control groups (n=497) was investigated. Logistic regression analysis, adjusting for age, pregnancy, oral contraceptive use, tobacco smoking and menopausal status, did not identify the SDF1­3' G801A polymorphism as a genetic risk factor for cervical cancer. The adjusted odds ratio (OR) for patients with the A/G, vs. G/G genotype was 1.203, with a 95% confidence interval (CI) of 0.909­1.591 (P=0.196). The adjusted OR for the A/A, vs. G/G genotype was 1.296 (95% CI=0.930­1.807; P=0.125) and for the A/A or A/G, vs. G/G genotype was 1.262 (95% CI=0.964­1.653; P=0.090)]. The P­value of the χ2 test of the trend observed for the SDF1­3' G801A polymorphism was at the borderline of being statistically significant (ptrend=0.0484). Stratified analyses between the distribution of the SDF1­3' G801A genotypes and cervical cancer risks demonstrated that this polymorphism may be a risk factor for patients with a positive history of tobacco smoking (1.778; 95% CI=1.078­2.934; P=0.0235). These findings suggested that the SDF1­3' G801A polymorphism may be a genetic risk factor for cervical cancer in patients with a positive history of tobacco smoking.

TNF-α -308 G/A as a risk marker of cervical cancer progression in the Polish population.

BACKGROUND AND OBJECTIVE: There are inconclusive data on the tumor necrosis factor-α (TNF-α) -308 G/A (rs1800629) polymorphism as a risk factor for cervical carcinogenesis. METHODS: Using high-resolution melting curve analysis, we investigated the prevalence of the TNF-α -308 G/A transition (rs1800629) in patients with cervical cancer (n = 362) and control subjects (n = 399). RESULTS: The p trend value calculated for the TNF-α -308 G/A transition was statistically significant (p trend = 0.026) for all patients. Logistic regression analysis with adjustment for age demonstrated that the A/A versus G/G genotype was significantly associated with cervical tumors. The adjusted odds ratio (OR) was 1.599 (95 % confidence interval [CI] 1.017-2.513; p = 0.042). Stratification of patients on the basis of the tumor stage revealed no contribution of the TNF-α -308 G/A transition to cervical cancer in stages I and II. However, we found a significant trend for the p value, as well as the contribution of TNF-α -308 G/A to cervical cancer, in stages III and IV. The p trend value was 0.003 in this group of patients. Moreover, logistic regression analysis with adjustment for age demonstrated that the adjusted OR for A/A versus G/G was 2.014 (95 % CI 1.122-3.613; p = 0.019) and the adjusted OR for A/A or A/G versus G/G was 1.583 (95 % CI 1.025-2.444; p = 0.038) for cervical cancer in stages III and IV. CONCLUSION: Our results indicate that the TNF-α -308 G/A transition is a risk factor for cervical cancer, particularly in stages III and IV.

Prognostic potential of DNA methylation and transcript levels of HIF1A and EPAS1 in colorectal cancer.

UNLABELLED: Hypoxic conditions during the formation of colorectal cancer may support the development of more aggressive tumors. Hypoxia-inducible factor (HIF) is a heterodimeric complex, composed of oxygen-induced HIFα and constitutively expressed HIFß subunits, which mediates the primary transcriptional response to hypoxic stress. Among HIFα isoforms, HIF1α (HIF1A) and endothelial PAS domain-containing protein 1 (EPAS1) are able to robustly activate hypoxia-responsive gene signatures. Although posttranslational regulation of HIFα subunits is well described, less is known about their transcriptional regulation. Here, molecular analysis determined that EPAS1 mRNA was significantly reduced in primary colonic adenocarcinoma specimens compared with histopathologically nonneoplastic tissue from 120 patients. In contrast, no difference in HIF1A mRNA levels was observed between cancerous and noncancerous tissue. Bisulfite DNA sequencing and high-resolution melting analysis identified significant DNA hypermethylation in the EPAS1 regulatory region from cancerous tissue compared with nonneoplastic tissue. Importantly, multivariate Cox regression analysis revealed a high HR for patients with cancer with low EPAS1 transcript levels (HR, 4.91; 95% confidence interval, CI, 0.42-56.15; P = 0.047) and hypermethylated EPAS1 DNA (HR, 33.94; 95% CI, 2.84-405.95; P = 0.0054). Treatment with a DNA methyltransferase inhibitor, 5-Aza-2'-deoxycytidine (5-aza-dC/Decitabine), upregulated EPAS1 expression in hypoxic colorectal cancer cells that were associated with DNA demethylation of the EPAS1 regulatory region. In summary, EPAS1 is transcriptionally regulated by DNA methylation in colorectal cancer. IMPLICATIONS: DNA methylation and mRNA status of EPAS1 have novel prognostic potential for colorectal cancer.