Liu Ping, Ma Chengbin, Wu Qiongwei, Zhang Wenying, Wang Cao, Yuan Li, Xi Xiaowei
1Gynecology Department, Shanghai General Hospital of Nanjing Medical University, Shanghai, 200080 China.
Gynecology Department, Changning Maternity and Infant Health Hospital, Shanghai, 200051 China.
Cancer Cell Int. 2019 Jul 11;19:178. doi: 10.1186/s12935-019-0897-8. eCollection 2019.
The aim of this study is to examine miRNA profiling and miR-369-3p participates in endometrioid adenocarcinoma (EEC) via the regulation of autophagy.
EEC and its adjacent normal tissues were obtained from 20 clinical patients after surgery. MiRNA profiling was performed using next generation sequencing (NGS) and was validated with quantitative real time PCR (qRT-PCR). qRT-PCR was also employed to measure miR-369-3p and autophagy-related protein 10 (ATG10) expression levels. Western blotting assay was performed to measure the expressions of ATG10 and LC3B. Luciferase reporter assay was conducted to confirm the direct targeting of ATG10 by miR-369-3p. Cell proliferation and migration assays were utilized to analyze the role of miR-369-3p in HEC-1-A cells.
We found that miR-369-3p expression levels were down-regulated in EEC compared to the control tissues. The overexpression of miR-369-3p inhibited cell proliferation and migration in EEC; furthermore, ATG10 expression increased in EEC tissues. ATG10 was found to be a potential target of miR-369-3p via a dual-luciferase reporter assay, and ATG10 was shown to be down-regulated by miR-369-3p in protein levels.
This study revealed that miR-369-3p inhibited cell proliferation and migration by targeting ATG10 via autophagy in EEC.
本研究旨在检测微小RNA（miRNA）表达谱，并探究miR-369-3p通过调控自噬参与子宫内膜样腺癌（EEC）的机制。
收集20例临床患者术后的EEC组织及其癌旁正常组织。采用新一代测序（NGS）技术进行miRNA表达谱检测，并通过定量实时荧光定量PCR（qRT-PCR）进行验证。同时，运用qRT-PCR检测miR-369-3p和自噬相关蛋白10（ATG10）的表达水平。采用蛋白质免疫印迹法检测ATG10和LC3B的表达。通过荧光素酶报告基因实验证实miR-369-3p对ATG10的直接靶向作用。利用细胞增殖和迁移实验分析miR-369-3p在HEC-1-A细胞中的作用。
我们发现，与对照组织相比，EEC中miR-369-3p的表达水平下调。miR-369-3p的过表达抑制了EEC细胞的增殖和迁移；此外，EEC组织中ATG10的表达增加。通过双荧光素酶报告基因实验发现ATG10是miR-369-3p的潜在靶点，并且miR-369-3p在蛋白质水平上可下调ATG10的表达。
本研究表明，在EEC中，miR-369-3p通过自噬靶向ATG10抑制细胞增殖和迁移。
