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*

摘要:

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。

Abstract:

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 long-effective 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.