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目的:探讨子宫内膜异位症(endometriosis,EMS)患者卵巢颗粒细胞中类固醇生成因子-1(steroidogenic factor-1,SF-1)表达下降的可能机制。方法:应用重亚硫酸盐测序法(bisulfite sequencing PCR,BSP)检测EMS患者和正常女性卵巢颗粒细胞SF-1基因启动子区5’Cp G位点甲基化水平,将测得的序列与原始序列比对,统计甲基化位点、数量并分析EMS组和正常组甲基化程度。结果:BSP片段涵盖SF-1基因转录起始位点附近的13个Cp G位点(从Cp G-84到Cp G+168),EMS组SF-1基因启动子区域的整体甲基化水平显著高于对照组(P<0.05),逐个位点的分析显示EMS组Cp G+7、+18、+21、+42、+54、+60、+132以及+146位点相较于对照组呈现高甲基化水平(P<0.05);而EMS组Cp G+77、+121和+141位点的高甲基化水平尤为显著(P<0.001)。结论:EMS患者卵巢颗粒细胞SF-1基因启动子区5’Cp G位点甲基化水平显著高于对照组,可能是EMS患者卵巢颗粒细胞SF-1 m RNA表达降低的机制。
Objective: To investigate the possible mechanism of decreased expression of steroidogenic factor-1 (SF-1) in ovarian granulosa cells in patients with endometriosis (EMS). Methods: Methylation level of 5’Cp G site in promoter region of SF-1 gene in ovarian granulosa cells of EMS patients and normal women was detected by bisulfite sequencing PCR (BSP). The measured sequence was compared with the original Sequence alignment, statistical methylation sites, the number and analysis of EMS group and normal group methylation level. Results: The BSP fragment covered 13 CpG sites (from Cp G-84 to CpG + 168) near the SF-1 transcriptional start site and the overall methylation level of SF-1 promoter region in EMS group The levels of CpG + 7, + 18, + 21, + 42, + 54, + 60, + 132 and +146 in EMS group were significantly higher than those in control group (P <0.05) (P <0.05), while the hypermethylation levels of CpG + 77, +121 and +141 in EMS group were more significant (P <0.001). Conclusion: The methylation level of 5’Cp G site of SF-1 gene in ovarian granulosa cells in EMS patients is significantly higher than that in control group, which may be the mechanism of decreased expression of SF-1 mRNA in ovarian granulosa cells of EMS patients.