腹腔镜子宫腺肌瘤切除术后辅助治疗对种植期内膜HOXA10及其基因启动子甲基化的影响

Impact of laparoscopic resection of adenomyoma combined with comprehensive assisted therapy on endometrial HOXA10 methylation during implantation window

目的探讨腹腔镜子宫腺肌瘤切除及术后促性腺激素释放激素激动剂(GnRH-a)、左炔诺孕酮宫内缓释系统(曼月乐,LNG-IUS)辅助治疗对内膜HOXA10-mRNA及其基因启动子区甲基化的影响。方法选取2013年6月-2016年6月就诊的子宫腺肌瘤77例,行腹腔镜子宫腺肌瘤切除术后,相继注射长效GnRH-a 6个月及放置曼月乐12个月,于术前及曼月乐取出后黄体中期取子宫内膜;同期已生育妇女24例作为对照组;RT-PCR法检测子宫内膜HOXA10-mRNA,亚硫酸氢盐修饰后测序(BSP)检测HOXA10基因启动子区F1片段甲基化状态。结果两组患者年龄无明显差异。观察组术前子宫内膜HOXA10-mRNA为(0.5±0.1),明显低于术后观察组(0.9±0.3)和对照组术前(0.9±0.4)。观察组术前子宫内膜HOXA10启动子F1片段甲基化位点数为(6.5±3.7),明显高于术后观察组(2.8±2.0)和对照组(2.2±1.8),差异有统计学意义(P<0.05)。结论子宫腺肌瘤降低内膜HOXA10的表达,腹腔镜子宫腺肌瘤切除术联合GnRH-a及曼月乐综合治疗,可降低内膜HOXA10启动子甲基化水平,提高HOXA 10-mRNA的表达,从而改善子宫内膜容受性。

Objective To observe the impact of laparoscopic adenomyoma resection combined with gonadotropin releasing hormone agonist(GnRH-a)and Levonorgestrel-Releasing Intrauterine System(LNG-IUS)on endometrial HOXA10 and HOXA10 promoter methylated CpG sites.Methods From June 2013 to June 2016,77 patients with adenomyoma received laparoscopic adenomyoma resection,after this operation they received longeffective GnRH-a for 6 months and LNG-IUS for 12 months respectively.Pipelle endometrial biopsies were obtained 7 days after urine luteal hormone surge before operation and removing LNG-IUS later.24 fertile patients were as control group and collected endometrium by the same method.Real-time polymerase chain reaction analyzed endometrial HOXA10-mRNA.HOXA10 promoter methylated CpG sites were tested by bisulfite salt modified sequencing.Results There was no significant difference about average age between the two groups.Endometrial HOXA10-mRNA of observation groups was(0.5±0.1)before operation,significantly less than that of observation groups after operation and control group.Endometrial HOXA10 promoter F1 fragment methylated CpG sites of observation groups were(6.5±3.7),significantly higher than that of observation groups after operation and control group.Conclusion Adenomyoma have determined expression of HOXA10-mRNA,resection of adenomyoma combined GnRH-a and LNG-IUS can increase endometrial HOXA10-mRNA expression,and reduce the methylated level of HOXA10 promoter during endometrial implantation.