摘要
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。
Ohjective： To gain further insight into the mechanism for differential steroidogenic factor- 1 （SF- 1） expression in normal and endometriotic granulosa cells. Methods： The methylation status of 5＇CpG sites of SF-1 promoter in normal and endometriotic granulosa cells were evaluated using bisulfite sequencing PCR. Results： Methylation status of a total of 13 CpG sites across a 213-bp region in the approximately 252-bp CpG sites （-84/＋186） at the SF-1 promoter was characterized by bisulfite genomic sequencing. Ten clones were checked for each involved subject. The 13 CpG sites of SF-1 were differentially methylated in normal group. However, increased methylation in endometriotic granulosa cells was observed when compared with normal granulosa cells （P〈0.05）. There found eleven significantly hypermethylated sites including CpG＋7,＋ 18,＋21,＋42, ＋54, ＋60, ＋77,＋121,＋ 132, ＋141 and ＋146 （P〈0.05）, in which CpG＋77, ＋121 and ＋141 were highly hypermethylated （P〈0.001）. Conclusion： We demonstrated that hypermethylation status of 5＇CpG site of SF-1 promoter in endometriotic granulosa cells, which may explain the mechanism for reduced SF-1 mRNA expression in granulosa cells of endometriosis.
出处
《生殖与避孕》
CAS
CSCD
北大核心
2016年第7期536-541,共6页
Reproduction and Contraception
基金
上海市自然科学基金资助项目
项目号:13ZR1445100
