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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

24卷

期数:

2018年06期

页码:

469-473

栏目:

基础医学

出版日期:

2018-11-20

文章信息/Info

Title:

Influence of miR-27a on apoptosis of gastric cancer cells in vitro

作者:

张 悦; 管叙文; 王婧雅; 黄鼎智

（天津医科大学肿瘤医院消化肿瘤内科，国家肿瘤临床医学研究中心，天津市“肿瘤防治”重点实验室，天津市恶性肿瘤临床医学研究中心，天津 300060）

Author(s):

ZHANG Yue;  GUAN Xu-wen;  WANG Jing-ya;  HUANG Ding-zhi

（Department of Gastrointestinal Medical Oncology, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin Key Laboratory of Cancer Prevention and Therapy, Tianjin’s Clinical Research Center for Cancer,Tianjin 300060, China）

关键词:

胃癌; miR-27a; FOXO1; 凋亡

Keywords:

gastric cancer;  miR-27a; FOXO1;  apoptosis

分类号:

R735.2

DOI:

-

文献标志码:

A

摘要:

目的：探讨miR-27a对胃癌SGC-7901细胞凋亡的影响及其作用机制。方法：分别构建miR-27a过表达和敲除FOXO1的SGC-7901细胞株，采用流式细胞术检测细胞凋亡情况。蛋白质印迹法（Western blot）检测细胞凋亡和自噬相关蛋白p53、BCL-2、LC3I、LC3II水平，同时检测上调及下调miRNA-27a的胃癌细胞FOXO1的表达水平。双荧光素酶报告基因分析法验证miR-27a的靶基因。实时聚合酶链式反应（Realtime PCR） 检测miR-27a高表达质粒、miR-27a低表达质粒的转染效率，并检测过表达/敲低miRNA-27a的胃癌细胞中FOXO1 mRNA的水平。结果：（1）上调miR-27a的细胞凋亡率较对照组明显下降，且凋亡相关蛋白BCL-2表达上调，p53水平下降，自噬相关蛋白LC3I、LC3II的表达较对照组下调（P<0.05）。而敲低胃癌细胞中FOXO1后得到了与上调miR-27a相似的实验结果。（2）双荧光素酶报告基因验证结果显示miR-27a与FOXO1存在靶点关系。miR-27a高表达质粒、miR-27a低表达质粒分别成功转染至胃癌细胞中。上调或下调胃癌细胞中miR-27a后，SGC-7901细胞中FOXO1 mRNA水平无明显变化（P>0.05），FOXO1基因表达在蛋白水平相应下调或上调。结论：体外实验中，miR-27a可通过靶向抑制FOXO1的表达抑制胃癌细胞凋亡，miR-27a/ FOXO1轴可以为胃癌治疗提供新的策略。

Abstract:

Objective: To explore the influence of miR-27a on apoptosis and mechanism in gastric cancer cell line SGC-7901. Methods：Gastric cancer (GC) cell lines SGC-7901 of over-expressed miR-27a and less-expressed FOXO1 were established. Flow cytometrywas used to measurecell apoptosis. The protein expressions of p53,BCL-2, LC3I and LC3II were detected by Western blot（WB）.The expressions of FOXO1 in cells of up-regulated and down-regulated miR-27a were also examined by WB. Dual luciferase reporterassay was used to verify the target gene of miR-27a. Real-time PCRwas employed to determine miR-27a levels of cells transfected by miR-27a mimics and inhibitors andlevels of FOXO1 mRNA. Results：Compared withnormal control cells, apoptosis rates of miR-27a up-regulated cells were decreased.WB suggested that BCL-2 expressions were up-regulated whilep53, LC3I and LC3II expressions were down-regulated as compared to control groups（P<0.05）. Experiments in cells of low-expressed FOXO1 had similar results.Results of dual luciferase reporter assay showed that FOXO1 was target gene of miR-27a．MiR-27a mimics and inhibitors were successfully transfected into GC cells. FOXO1 mRNA levels had no significant change in cells transfected by miR-27a mimics and inhibitors（P>0.05）. MiR-27a negatively regulated FOXO1 protein expression. Conclusion：In vitro, miR-27a inhibits GC cell apoptosis by targeting FOXO1. miR-27a/ FOXO1 axis maybe a new therapeutic target in gastric cancer.

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备注/Memo

备注/Memo:

基金项目 国家自然科学基金资助项目（81572321）；天津市应用基础与前沿技术研究计划（15JCYBJC28200）
作者简介 张悦（1991-），女，硕士在读，研究方向：肿瘤学；通信作者：黄鼎智，E-mail：hdz199109@163.com。

常用功能

更新日期/Last Update: 2018-11-30