摘要： 目的 研究微小RNA-34a-5p（miR-34a-5p）靶向AKT1基因对子宫内膜异位症（EM）子宫内膜基质细胞（ESCs）侵袭及自噬的调控作用。方法 原代分离、培养EM患者ESCs，构建negative control RNA（NC）及mimic细胞模型，采用双荧光肾素酶报告系统验证miR-34a-5p与AKT1基因存在结合位点，RT-PCR及Western blotting检测并验证2组细胞miR-34a-5p和AKT1基因的表达及调控关系；检测miR-34a-5p对ESCs的增殖、迁移、侵袭、凋亡的影响。结果 EM患者ESCs中miR-34a-5p较非EM患者明显降低，而AKT1基因的表达明显升高，且两者变化呈负相关；miR-34a-5p通过特异性结合AKT1基因的3'UTR区对AKT1基因的表达进行调控；转染miR-34a-5p mimic可使ESCs中miR-34a-5p表达升高，AKT1基因表达降低，同时使ESCs的增殖、迁移及侵袭能力增强，而凋亡及自噬能力下降。结论 miR-34a-5p靶向AKT1基因促进ESCs的增殖、迁移及侵袭能力，降低其凋亡及自噬能力，可能在EM的发病过程中发挥作用。

miR-34a-5p靶向AKT1基因对子宫内膜异位症子宫内膜基质细胞侵袭及自噬的调控

刘敏娟^1,2, 邓月秀³, 马颖¹

1. 南方医科大学珠江医院妇产科, 广州 510282;
2. 东莞市人民医院妇产科, 广东 东莞 523000;
3. 桂林市妇幼保健院妇产科, 广西 桂林 541001

收稿日期:2019-09-19出版日期:2020-09-30发布日期:2020-09-15
通讯作者:马颖E-mail:mayingwuzhuoyi@126.com
作者简介:刘敏娟(1993-),女,硕士研究生.
基金资助:国家自然科学基金（81701418）；广东省科技计划（2014A020212667）


关键词: 子宫内膜异位症, 微小RNA-34a-5p, AKT1基因, 子宫内膜基质细胞
Abstract: Objective To investigate the effect of microRNA-34a-5p (miR-34a-5p) on the invasion and autophagy of endometrial stromal cells (ESCs) during endometriosis (EM). Methods ESCs were isolated from EM patients and the expression of miR-34a-5p and AKT1 in ESCs were analyzed by RT-PCR and the expression of AKT1 protein was detected by western blotting. A dual-luciferase reporter assay was used to dissect the interaction between miR-34a-5p and AKT1. The proliferation,migration,invasion,and apoptosis of ESCs transfected with miR-34a-5p mimic or negative control (NC) was measured with the CCK-8 assay,wound-healing assay,Transwell-invasion assay,and flow cytometry,respectively. Results During EM,the expression of miR-34a-5p in the ESCs was significantly lower than that in non-EM patients (P<0.05),while AKT1 expression was significantly higher. miR-34a-5p could specifically bind the 3'-UTR of AKT1 and regulate its expression. The expression of AKT1 in the ESCs transfected with the miR-34a-5p mimic was lower than in the NC group. Thus,overexpression of miR-34a-5p can reduce the proliferation,migration,and invasion of ESCs in EM and enforce apoptosis. Conclusion miR-34a-5p promotes the viability of ESCs by targeting AKT1,which may play a crucial role in the pathogenesis of EM.
Key words: endometriosis, microRNA-34a-5p, AKT1 gene, endometrial stromal cell
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