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微小RNA-101-3p通过靶向EZH2诱导子宫内膜癌细胞自噬。

miR-101-3p induces autophagy in endometrial carcinoma cells by targeting EZH2.

作者信息

Wang Cuilan, Liu Bo

机构信息

Department of Gynaecology and Obstetrics, Jinan Maternity and Child Care Hospital, No. 2 Xiaojingsan Road, Jinan, 250001, Shandong, China.

出版信息

Arch Gynecol Obstet. 2018 Jun;297(6):1539-1548. doi: 10.1007/s00404-018-4768-7. Epub 2018 Apr 24.

DOI:10.1007/s00404-018-4768-7
PMID:29691644
Abstract

OBJECTIVE

This study aimed to investigate the effect of miR-101-3p on autophagy in endometrial carcinoma (EC) cells and the connection between miR-101-3p and EZH2.

METHODS

The expression levels of miRNAs were analyzed by microarray. The expression level of autophagy related proteins was measured by western blot. The mRNA expression level of beclin-1 was determined by qRT-PCR. Autophagy in EC cells was traced by GFP-LC3 fusion protein and observed by fluorescence microscopy. The number of autophagic vacuoles was determined by transmission electron microscopy (TEM). A luciferase reporter assay was utilized to assess the target relationship between miR-101-3p and EZH2.

RESULTS

The expression level of miR-101-3p in EC tissues was lower than in normal tissues. miR-101-3p upregulated the expression levels of the autophagy-related proteins LC3-II and beclin-1 in EC cells in a time- and dose-dependent manner. Overexpression of miR-101-3p and silencing of EZH2 both promoted autophagy in EC cells. Luciferase reporter assays verified that miR-101-3p inhibited EZH2 expression by binding to its 3'-UTR region.

CONCLUSION

miR-101-3p promoted autophagy in EC cells by downregulating the expression of EZH2, and it induced autophagy in EC cells by suppressing EZH2 expression. Inhibition of miR-101-3p could reduce its autophagy induction effect on EC cells.

摘要

目的

本研究旨在探讨miR-101-3p对子宫内膜癌（EC）细胞自噬的影响以及miR-101-3p与EZH2之间的联系。

方法

通过基因芯片分析miRNA的表达水平。采用蛋白质免疫印迹法检测自噬相关蛋白的表达水平。通过qRT-PCR测定beclin-1的mRNA表达水平。利用GFP-LC3融合蛋白追踪EC细胞中的自噬，并通过荧光显微镜观察。通过透射电子显微镜（TEM）确定自噬泡的数量。利用荧光素酶报告基因检测法评估miR-101-3p与EZH2之间的靶向关系。

结果

EC组织中miR-101-3p的表达水平低于正常组织。miR-101-3p以时间和剂量依赖性方式上调EC细胞中自噬相关蛋白LC3-II和beclin-1的表达水平。miR-101-3p的过表达和EZH2的沉默均促进了EC细胞中的自噬。荧光素酶报告基因检测证实miR-101-3p通过与其3'-UTR区域结合抑制EZH2表达。

结论

miR-101-3p通过下调EZH2的表达促进EC细胞中的自噬，并且它通过抑制EZH2表达诱导EC细胞中的自噬。抑制miR-101-3p可降低其对EC细胞的自噬诱导作用。

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