摘要
目的:探讨miR-181d-5p通过与人第10号染色体缺失的磷酸酶和张力蛋白同源的基因(PTEN)mRNA 3'UTR区相互作用,参与小鼠睾丸支持细胞TM4细胞间紧密连接损伤及其可能机制。方法:通过网站预测miR-181d-5p的靶基因为PTEN mRNA,采用双荧光素酶报告基因分析实验在293T细胞上验证其与靶基因3'UTR区的结合;在TM4细胞上采用mimic过表达miR-181d-5p,引起PTEN蛋白表达降低,再进行p-FAK-Tyr397、p-Akt-Thr308等相关信号分子检测以及细胞间跨膜电阻(TER)值的测定。结果:MiR-181d-5p能与PTEN mRNA的3'UTR区结合,并降低其表达;在TM4细胞上过表达miR-181d-5p后,致使p-FAK-Tyr397和p-Akt-Thr308水平升高,以及TER值降低。结论:MiR-181d-5p通过与PTEN mRNA 3'UTR区结合降低其表达,引起p-FAK-Tyr397水平升高,以及PI3K/Akt信号通路激活,导致睾丸支持细胞间紧密连接损伤。
Objective:To investigate if miR-181d-5p can combine with the 3'UTR of PTEN mRNA and participate in the tight junction damage between mouse testis Sertoli cells(TM4 cell line),as well as the underlying mechanism.Methods:The target gene of miR-181d-5p was predicted via the website,and the dual-luciferase reporter system was used to verify the combination of miR-181d-5p and PTEN mRNA in 293T cell line;the miR-181d-5p was over-expressed using mimic in TM4 cell line,and then the level of signaling molecules(e.g.p-FAK-Tyr397 and p-Akt-Thr308)and trans-epithelium electrical resistant(TER)value were detected.Results:The result showed that miR-181d-5p could combine with the 3'UTR of PTEN mRNA and down-regulate the expression of PTEN;in addition,the over-expression of miR-181d-5p in TM4 cell line resulted in the decrease of TER value and the up-regulation of p-FAK-Tyr397 and p-Akt-Thr308.Conclusion:MiR-181d-5p decreases the the expression of PTEN through interacting with the 3'UTR of PTEN mRNA,then brings the up-regulation of p-FAK-Tyr397 and activation of PI3K/Akt signaling pathway,finally contributing to the disruption of tight junction between mouse testis Sertoli cells.
作者
张丽虹
王锋
赵小贞
王玮
Zhang Lihong;Wang Feng;Zhao Xiaozhen;Wang Wei(Department of Human Anatomy,Laboratory of Clinical Applied Anatomy,Key Laboratory of Aging and Neurodegenerative Disease,Fujian Medical University,Fuzhou 350122,China)
出处
《解剖学杂志》
CAS
2021年第2期95-100,共6页
Chinese Journal of Anatomy
基金
福建省卫生计生科研人才培养项目(2018-1-74)
福建医科大学启航基金(2017XQ1005)
福建医科大学高层次人才科研启动经费项目(XRCZX2017033)
福建省自然科学基金(2017J01819)
国家卫生计生委共建科研基金-卫教联合攻关(WKJ2016-2-35)。