miR-145通过调控人子宫内膜基质细胞OCT4的表达促进子宫内膜异位症的发展机制  

miR-145 Regulates the Expression of OCT4 in Human Endometrial Stromal Cells and Promotes the Development of Endometriosis

作　　者：张素仙[1] 冯莉嵋 伍路 杨绍艳 赵庆华[1] ZHANG Suxian;FENG Limei;WU Lu;YANG Shaoyan;ZHAO Qinghua(Dept.of Gynecology,The Second Affiliated Hospital of Kunming Medical University,Kunming Yunnan 650101,China;Dept.of Pharmacy,The Second Affiliated Hospital of Kunming Medical University,Kunming Yunnan 650101,China)

机构地区：[1]昆明医科大学第二附属医院妇科,云南昆明650101 [2]昆明医科大学第二附属医院药学部,云南昆明650101

出　　处：《昆明医科大学学报》2022年第2期23-33,共11页Journal of Kunming Medical University

基　　金：云南省教育厅科学研究基金资助项目(2020J0183);云南省科学技术厅-昆明医科大学应用基础研究联合专项基金资助(202001AY070001-227);昆明医科大学第二附属医院院内科研项目(2018yk017)。

摘　　要：目的通过体外细胞试验探讨miR-145在子宫内膜异位症(Endometriosis,EMs)中的调控机制。方法蛋白印迹检测OCT4等相关蛋白的表达水平,通过相关分析和双荧光素酶报告试验用于评估miR-145和OCT4之间的关联,CCK8试剂盒检测细胞活力,流式细胞术检测细胞凋亡,通过Transwell法检测hESCs的迁移。结果前期临床试验发现异位子宫内膜组织miR-145表达上调,OCT4表达下调。在细胞实验中,miR145过表达可显著促进hESCs的增殖和迁移(P<0.01),但抑制hESCs的凋亡(P<0.05)。miR-145模拟物转染hESCs后,OCT4、Bax和MMP1等蛋白表达水平降低(P<0.01),Bcl-2蛋白表达水平升高(P<0.05)。敲除miR145逆转了上述结果,并通过靶向OCT4显著抑制了hESCs的增殖(P<0.05)。结论研究结果表明,miR-145表达增加通过抑制OCT4蛋白表达,可能通过促进子宫内膜基质细胞上皮细胞间质转化(Epithelial-mesenchymal Transition,EMT)过程,从而在促进EMs的发展中起到一定的作用。Objective To further explore the regulatory mechanism of miR-145 in Endometriosis(EMs)through cell experiments.Methods miR-145 or octamer-binding transcription factor 4(OCT4)gene and protein expression levels in cells were examined using reverse transcription-quantitative PCR and western blotting,respectively.Correlation analyses and dualluciferase reporter assays were performed to assess the association between miR-145 and OCT4.A Cell Counting Kit-8 assay was performed to test cell viability,while cell apoptosis was measured using flow cytometry.Moreover,the migration of hESCs was measured via Transwell assays.Results In cell assays,overexpression of miR-145 signifcantly promoted proliferation and migration(P<0.01),but inhibited the apoptosis of hESCs(P<0.05).Furthermore,the transfection of hESCs with miR-145 mimics decreased the protein expression levels of OCT4,Bax and MMP1(P<0.01),as well as increased the protein expression of Bcl2.However,knockdown of miR-145 reversed these results and significantly inhibited the proliferation of hESCs by targeting OCT4(P<0.05).Conclusion The present results suggested that knockdown of miR-145 signifcantly suppressed the development of EMs by targeting OCT4,which may serve as a potential target for the treatment of EMs.

关 键 词：子宫内膜异位症 miRNA-145 OCT4 EMT 人子宫内膜基质细胞 

分 类 号：R711.71[医药卫生—妇产科学]