miR-328-5p通过调控RAGE对异位子宫内膜细胞生物学功能的影响  

Effects of miR-328-5p on the biological function of ectopic endometrial cells by regulating RAGE

作　　者：韩奇倚 赵楚楚 HAN Qiyi;ZHAO Chuchu(Lishui Municipal Central Hospital,Lishui,Zhejiang 323000,China)

机构地区：[1]丽水市中心医院,浙江丽水323000

出　　处：《中国优生与遗传杂志》2023年第4期726-732,共7页Chinese Journal of Birth Health & Heredity

基　　金：浙江省医药卫生科技计划项目(2021KY1243);丽水市科技计划项目(2019SJZC28)。

摘　　要：目的 探讨miR-328-5p在异位子宫内膜细胞生物学功能中的作用及潜在机制。方法 实时荧光定量聚合酶链反应(RT-qPCR)和Western blot检测异位子宫内膜组织中miR-328-5p、RAGE mRNA和蛋白的表达。分离并培养异位子宫内膜基质细胞,将其分为NCmimic组、miR-328-5pmimic组、miR-328-5pmimic+Vector组、miR-328-5p mimic+pcDNA-RAGE组,RT-qPCR检测细胞中miR-328-5p和RAGE mRNA的表达;CCK-8实验检测细胞增殖能力;Transwell实验检测细胞侵袭和迁移能力;Westernblot实验检测RAGE蛋白的表达;生物信息学和双荧光素酶报告基因实验用来预测和验证miR-328-5p和RAGE的靶向关系。结果 与正常子宫内膜组织比较,异位子宫内膜组织中miR-328-5p表达显著降低,RAGE mRNA和蛋白表达显著升高(P<0.05)。与NC mimic组比较,miR-328-5p mimic组细胞中miR-328-5p表达显著升高,RAGE mRNA和蛋白表达显著降低,细胞增殖、侵袭和迁移能力显著降低(P<0.05)。与NC mimic组比较,WT-RAGE和miR-328-5p mimic共转染的细胞荧光素酶活性显著降低(P<0.05),而MUT-RAGE和miR-328-5pmimic共转染的细胞的荧光素酶活性无显著性差异(P>0.05)。与miR-328-5pmimic+Vector组比较,miR-328-5pmimic+pcDNA-RAGE组细胞增殖、侵袭和迁移能力显著升高(P<0.05)。结论 过表达miR-328-5p通过调控RAGE抑制异位子宫内膜基质细胞增殖、迁移和侵袭能力。Objective To explore the role and underlying mechanism of miR-328-5p in the biological function of ectopic endometrial cells.Methods Real-time quantitative polymerase chain reaction(RT-qPCR)and Western blot were used to detect the mRNA and protein expressions of miR-328-5p and RAGE in ectopic endometrial tissue.The ectopic endometrial stromal cells were isolated and cultured,and divided into NC mimic group,miR-328-5p mimic group,miR-328-5p mimic+Vector group and miR-328-5p mimic+pcDNA-RAGE group.RT-qPCR was used to detect the expression of miR-328-5p and RAGE mRNA.CCK-8 assay was used to detect the cell proliferation.Transwell assay was used to detect cell invasion and migration.Western blot was used to detect the expression of RAGE protein.The bioinformatics and dual luciferase reporter gene experiments were used to predict and validate the targeting relationship between miR-328-5p and RAGE.Results Compared with normal endometrial tissue,the expression of miR-328-5p in ectopic endometrial tissue was significantly decreased,while the mRNA and protein expressions of RAGE were significantly increased(P<0.05).Compared with the NC mimic group,the expression of miR-328-5p was significantly increased,the mRNA and protein expressions of RAGE were significantly decreased,and the ability of cell proliferation,invasion and migration were significantly decreased in the miR-328-5p mimic group(P<0.05).Compared with NC mimic group,the luciferase activity of the cells cotransfected with WT-RAGE and miR-328-5p mimic was significantly decreased(P<0.05),but the luciferase activity of the cells cotransfected with MUT-RAGE and miR-328-5p mimic had no significant difference(P>0.05).Compared with the miR-328-5p mimic+Vector group,the ability of cell proliferation,invasion and migration were significantly increased in the miR-328-5p mimic+pcDNA-RAGE group(P<0.05).Conclusion Overexpression of miR-328-5p could inhibit the proliferation,migration and invasion of ectopic endometrial stromal cells by regulating RAGE expression.

关 键 词：miR-328-5p RAGE 子宫内膜异位症 增殖 侵袭 

分 类 号：R711.71[医药卫生—妇产科学]