吉林大学学报(医学版) ›› 2015, Vol. 41 ›› Issue (02): 240-244.doi: 10.13481/j.1671-587x.20150207

• 基础研究 • 上一篇    下一篇

miR-18a对结直肠癌SW116细胞辐射敏感性的影响

燕蒙蒙1, 徐珊1, 高岩2, 刘扬1, 贺梦子1, 陈司霖1, 李鹏武1, 刘晓冬1, 马淑梅1   

  1. 1. 吉林大学公共卫生学院 卫生部放射生物学重点实验室, 吉林 长春 130021;
    2. 吉林大学第一医院肿瘤中心放疗科, 吉林 长春 130021
  • 收稿日期:2014-07-17 出版日期:2015-03-28 发布日期:2015-04-04
  • 通讯作者: 马淑梅, 教授, 硕士研究生导师(Tel:0431-85619443, E-mail:shmm2001@126.com) E-mail:shmm2001@126.com
  • 作者简介:燕蒙蒙(1988-), 女, 山东省东营市人, 在读医学硕士, 主要从事肿瘤的放射生物学方面的研究。
  • 基金资助:

    国家自然科学基金资助课题(30970682);吉林省科技厅科研基金资助课题(201205010)

Effect of miR-18a on irradiation sensitivity of colorectal cancer SW116 cells

YAN Mengmeng1, XU Shan1, GAO Yan2, LIU Yang1, HE Mengzi1, CHEN Silin1, LI Pengwu1, LIU Xiaodong1, MA Shumei1   

  1. 1. Key Laboratory of Radiobiology, Ministry of Health, School of Public Health, Jilin University, Changchun 130021, China;
    2. Department of Radiotherapy, Tumor Center, First Hospital, Jilin University, Changchun 130021, China
  • Received:2014-07-17 Online:2015-03-28 Published:2015-04-04

摘要:

目的:探讨miR-18a与结直肠癌SW116细胞辐射敏感性之间的关系,阐明miR-18a影响细胞凋亡和自噬的可能机制。方法:人结直肠癌SW116细胞分为miR-18a NC组、miR-18a mimic组、miR-18a NC+4Gy组和miR-18a mimic+4Gy组。qRT-PCR法检测照射前后SW116细胞中miR-18a的表达;集落形成实验观察miR-18a对SW116细胞辐射敏感性的影响;GFPLC3形态学方法检测SW116细胞自噬率;流式细胞术检测SW116细胞凋亡率;采用生物信息学方法预测miR-18a的靶基因,以荧光素酶报告基因验证miR-18a与靶基因3'UTR结合;Western blotting法检测SW116细胞中共济失调-毛细血管扩张突变基因(ATM)蛋白表达。结果:与照射前比较,照射后SW116细胞中miR-18a表达水平明显降低(P<0.05)。集落形成实验,与miR-18a NC组比较,miR-18a mimic组SW116细胞辐射敏感性增强(P<0.05),细胞凋亡率和自噬率明显增加(P<0.05)。与miR-18a NC+4Gy组比较,miR-18a mimic+4Gy组SW116细胞中ATM蛋白表达减少。结论:miR-18a的靶基因为ATM;miR-18a mimic可促进辐射诱导的细胞凋亡和自噬,且能增加结直肠癌SW116细胞辐射敏感性。

关键词: miR-18a, 结直肠肿瘤, 辐射敏感性, 细胞凋亡, 自噬

Abstract:

Objective To discuss the relationship between miR-18a and the irradiation sensitivity of the colorectal cancer SW116 cells,and to elucidate the possible mechanism of the effects of miR-18a on the apoptosis and autophagy of the cells.Methods The SW116 cells were divided into miR-18a NC group,miR-18a mimicgroup, miR-18a NC+4Gy group,and miR-18a mimic+4Gy group. The expression of miR-18a in SW116 cells was detected by qRT-PCR before and after irradition;colony-forming assay was used to detect the effect of miR-18a on the irradiation sensitivity of the SW116 cells;GFPLC3 morphological assay was used to detect the autophagy rate of SW116 cells;the apoptotic rate of SW116 cells was analyzed by flow cytometry;the target gene of miR-18a was predicted by bioinformatics methods and dual-luciferase reporter assay was used to demonstrate the binding of miR-18a and 3'UTR targets;the expression of ataxia-telangiectasia mutated(ATM) protein in SW116 cells was measured by Western blotting method.Results compared with before irradation,the expression level of miR-18a in the SW116 cells after irradation was decreased obviously (P<0.05). The colony-forming assay results showed that the irradiation sensitivity of the SW116 cells in miR-18a mimic group was increased compared with miR-18a NC group(P<0.05),and the apoptotic rate and autophagy rate of the SW116 cells in miR-18a mimic group were increased (P<0.05). Compared with miR-18a NC+4Gy group,the expression level of ATM protein in miR-18a mimic+4Gy group was decreased.Conclusion ATM is the target gene of miR-18a in the SW116 cells. miR-18a mimic can increase the apoptosis and autophagy induced by irradition,and can increase the radiosensitivity of colorectal cancer SW116 cells.

Key words: miR-18a, colorectal neoplasms, radiation sensitivity, apoptosis, autophagy

中图分类号: 

  • R735.35