| Objective: To explore the association of single nucleotide polymorphism(SNP) in micro RNA coding regions and binding sites with pelvic organ prolapse(POP) risk, as well as the interactions with genetic variants and some environmental factors with POP risk, which aims to provide scientific basis for the diagnosis, treatment, and prevention of POP.Methods: From March 2012 to March 2013, 291 cases diagnosed with POP were enrolled, and 300 healthy controls matched with cases by age and resident areas were recruited in the same periods, all the participants came to gynaecology and obstetrics department of Fuzhou General Hospital of Nanjing Military Command. Peripheral venous blood(2ml) was drawn from all participants, and the vaginal wall tissues were only from the patients who need surgery. Genomic DNA was extracted from peripheral blood. Single nucleotide polymorphisms in mi R-196a-2(rs11614913), mi R-146a(rs2910164), mi R-149(rs2292832), mi R-499(rs3746444) encoding regions and 3’untranslated region(3’UTR) of COL1A1(rs1061947), COL1A1(rs1061237), MMP1(rs2071230), MMP1(rs5854), MMP1(rs470215), MMP2(rs7201), LOXL1(rs3522), MMP9(rs9509), MMP9(rs1056628), were selected and their biological functions were evaluateded with the relevant literatures and bioinformatics software. The Sequenom Mass ARRAY System was used to detect the genotypes of the SNPs. The SPSS statistical software package version 17.0 was used for all the statistical analyses, all tests were 2-sided and P<0.05 considered statistically significant. The Hardy-Weinberg equilibrium(HWE) and genotype frequencies difference betweenPOP group and the controls were using χ2-test, whereas the genetic risk in POP patients was estimated by logistic regression analysis, and it was expressed as odds ratios(ORs) with 95% confidence intervals(CIs). And stratified analysis was used to analyse the interactions between the genetic variants and environmental factors. The expression of COL1A1 and MMP1 m RNA in vaginal wall tissues were detected by Real time-PCR technique, in transcript levels; The levels of COL1A1 and MMP1 protein in vaginal wall tissues were detected by Western blotting, in translate levels.Results: 1. SNPs within mi R-196a-2(CC vs TT) encoding region was associated with increased POP risk(OR=2.00, 95%CI:1.19-3.36, P=0.009), and after stratification by environmental factors, significant associations between the SNP and POP risk was observed among those with the menopausal status(CC vs TT, OR=2.13, 95%CI: 1.11-4.08, P=0.024 and CC/CT vs TT, OR=1.60, 95%CI:1.01-2.55, P=0.047), one or more than one vaginal deliveries history(OR=2.26, 95%CI:1.10-4.63, P=0.026), and body mass index(BMI) ≥23.5(CC vs TT, OR=2.48, 95%CI:1.16-5.32, P=0.019 and CC/CT vs TT, OR=2.00, 95%CI:1.14-3.51, P=0.016). For the SNP within mi R-146 a, no significant association was observed among the two groups, however significant association was detected between the SNP(CC vs TT) and reduced POP risk among those with pre-menopause(OR=0.09, 95%CI:0.01-0.81, P=0.031). For the SNP within mi R-499, no significant association was observed among the two groups, however significant association was detected between the SNP(CC/CT vs TT) and POP risk among those with BMI≥23.5(OR=1.77, 95%CI:1.03-3.04, P=0.038). 2. The SNP(rs1061237) in 3’UTR of COL1A1 was associated with reduced POP risk(OR=0.59, 95%CI:0.37-0.95, P=0.031) and after stratification by environmental factors, significant associations between the SNP and reduced POP risk was observed among those with no history of vaginal delivery(TT vs CC, OR=0.38, 95%CI: 0.18-0.79, P=0.009 and CT/TT vs CC, OR=0.48, 95%CI:0.27-0.87, P=0.016). The SNPs(rs470215 and rs5854) in 3’UTR of MMP1 were associated with increased POPrisk(OR=2.01, 95%CI:1.16-3.51, P=0.013 and OR=2.76, 95%CI:1.20-6.38, P=0.017 respectively). And after stratification by environmental factors, significant associations between the rs5854 and POP risk was observed among those with the menopausal status(CT/TT vs CC, OR=2.05, 95%CI:1.08-3.87, P=0.028 and TT vs CC, OR=3.37, 95%CI:1.27-8.94, P=0.015), one or more than one vaginal deliveries history(CT/TT vs CC, OR=2.21, 95%CI: 1.12-4.36, P=0.022 and OR=3.95, 95%CI: 1.43-10.79, P=0.007), and BMI≥23.5(CT/TT vs CC, OR=2.44, 95%CI:1.06-5.61, P=0.036 and CC/CT vs TT, OR=4.43, 95%CI:1.21-16.18, P=0.024). For the SNP(rs2071230) within in 3’UTR of MMP1, no significant association was observed among the two groups, however significant association was detected between the SNP(GA/GG vs AA) and reduced POP risk among those with pre-menopause(OR=0.37, 95%CI:0.19-0.72, P=0.003). 3. There was no significant difference among the three groups on the m RNA expression of COL1A1(rs1061237). However, the protein expression was higher in genotype TT than genotype CT or CC, whereas the protein expression was higher in genotype CT than genotype CC. 4. There was no significant difference among the three groups on the m RNA expression of MMP1(rs5854). However, the protein expression was higher in genotype TT than genotype CT or CC, whereas the protein expression was higher in genotype CT than genotype CC.Conclision: 1. The SNP in mi R-196a-2 was associated with increased POP risk, and this SNP might be able to interact with the menopausal status, the vaginal deliveries history and BMI≥23.5; SNP within mi R-146 a could interact with menopausal status to modify POP risk. SNP within mi R-499 could interact with BMI to modify POP risk. 2. The SNP(rs1061237) in 3’UTR of COL1A1 was associated with increased POPrisk, and the SNP might be able to interact with vaginal delivery. The SNP(rs5854) in 3’UTR of MMP1 was associated with increased POP risk, and the SNP might be able to interact with the menopausal status, vaginal delivery history and BMI≥23.5. SNP(rs470215) in 3’UTR of MMP1 was associated with increased POP risk. SNP(rs2071230) in 3’UTR of MMP1 could be able to interact with the menopausal status to modify POP risk. 3. The polymorphism in COL1A1(rs1061237) could participate the pathogenesis of POP by affecting the ability of mi R-3119 combining to the target sites in COL1A1. 4. The polymorphism in MMP1(rs5854) could participate the pathogenesis of POP by affecting the ability of mi RNA combining to the target sites in 3’UTR of MMP1. |
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