miR-34a-5p及AKT1基因在子宫内膜异位症子宫内膜组织中的表达及其对子宫内膜基质细胞迁移和侵袭的影响  

Expression of miR-34a-5p and AKT1 gene in endometrial tissue of endometriosis and its effect on migration and invasion of endometrial stromal cells

作　　者：冯婉琴 邓月秀 马颖[1] Feng Wan-Qin;Deng Yue-Xiu;Ma Ying(Department of Gynaecology and Obstetrics,Zhujiang Hospital of Southern Medical University,Guangzhou 510282,China;Department of Gynaecology and Obstetrics,Maternal and Child Health Center in Guilin City,Guilin,Guangxi 541001,China)

机构地区：[1]南方医科大学珠江医院妇产科,广州510282 [2]桂林市妇幼保健院妇产科,广西桂林541001

出　　处：《解放军医学杂志》2020年第5期503-508,共6页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(81701418);广东省省级科技计划项目(2014A020212667)。

摘　　要：目的探讨miR-34a-5p及AKT1基因在子宫内膜异位症(EM)患者子宫内膜组织中的表达及其对子宫内膜基质细胞(ESC)迁移和侵袭的影响。方法收集2018年1月-2019年6月因良性妇科疾病在南方医科大学珠江医院妇产科行子宫切除术的患者91例,分为EM组(n=68)与非EM组(n=23),获取其子宫内膜组织,采用原位杂交及免疫组化法检测miR-34a-5p及AKT1基因在子宫内膜组织中的表达情况;使用脂质体-3000将miR-34a-5p mimic和阴性对照RNA(miR-NC)对ESC细胞进行转染,构建miR-34a-5p mimic组及miR-NC组细胞模型,培养12、24、48 h后,采用CCK-8法检测细胞增殖率,采用细胞迁移、侵袭、凋亡及自噬能力实验检测miR-34a-5p对ESC增殖、迁移、侵袭、凋亡及自噬的影响。结果EM组子宫内膜组织中的miR-34a-5p阳性表达率低于非EM组(16.2%vs.82.6%,χ~2=34.323,P<0.001),AKT1阳性表达率高于非EM组(72.1%vs.30.4%,χ~2=12.581,P<0.001)。miR-NC组培养12、24、48 h后的细胞增殖率均高于miR-34a-5p mimic组,差异有统计学意义(P<0.05);miR-NC组细胞迁移能力(65.00%±5.00%)高于miR-34a-5p mimic组(30.67%±4.04%),差异有统计学意义(t=9.25,P<0.05);miR-NC组的侵袭能力较miR-34a-5p mimic组明显增强[(88.0±8.5)个vs.(32.3±6.1)个,t=9.179,P<0.05];miR-NC组的细胞凋亡率(9.33%±3.51%)明显低于miR-34a-5p mimic组(18.00%±2.00%);miR-NC组中LC3基因的表达水平明显低于miR-34a-5p mimic组[(0.19±0.04)vs.(0.39±0.03),t=8.02,P<0.05]。结论miR-34a-5p可能通过靶向AKT1基因来影响ESC的增殖、迁移、侵袭、凋亡及自噬功能,从而参与EM的发病。Objective To investigate the expression of miR-34a-5p and AKT1 genes in endometrium tissues of patients with endometriosis (EM) and their effects on migration and invasion of endometrial stromal cells (ESCs).Methods A total of 91 patients,undergone hysterectomy in the Department of Obstetrics and Gynecology of Zhujiang Hospital of Southern Medical University from Jan.2018 to Jun.2019 due to benign gynecological diseases,were collected and divided into EM group (68 cases) and non-EM group (23 cases).The expressions of miR-34a-5p and AKT1 genes in endometrium tissues of patients were detected by in situ hybridization and immunohistochemistry.ESCs were transfected with miR-34a-5p mimic and negative control RNA (miR-NC) using liposome-3000 to construct the cell models of miR-34a-5p mimic group and miR-NC group.The cell proliferation rate was detected by CCK-8 method,and cell migration,invasion,apoptosis and autophagy ability experiments were performed to determine the effect of miR-34a-5p on ESCs’ proliferation,migration,invasion,apoptosis and autophagy.Results The positive expression rate of miR-34a-5p was lower,and of AKT1 was higher in EM group than those in non-EM group (16.2% vs.82.6%,χ~2=34.323;72.1% vs.30.4%,χ~2=12.581,P<0.001).After culturing for 12,24 and 48 h,the cell proliferation rate was higher in miR-NC group than that in miR-34a-5p mimic group (P<0.05).The cell migration ability and invasion ability were higher in miR-NC group than those in miR-34a-5p mimic group with statistically significant difference [(65.00%±5.00%) vs.(30.67%±4.04%);(88.0±8.5) vs.(32.3±6.1),t=9.179,P<0.05].The cell apoptosis rate and the expression level of LC3 gene were obviously lower in miR-NC group than those in miR-34a-5p mimic group [(9.33%±3.51%) vs.(18.00%±2.00%);(0.19±0.04) vs.(0.39±0.03),t=8.02,P<0.05].Conclusion miR-34a-5p may be involved in the pathogenesis of EM by targeting AKT1 genes to affect the proliferation,migration,invasion,apoptosis and autophagy function of ESC.

关 键 词：子宫内膜异位症 miR-34a-5p AKT1基因 免疫组化 原位杂交 迁移 侵袭 

分 类 号：R71[医药卫生—妇产科学]