miR-18a对结直肠癌SW116细胞辐射敏感性的影响

doi:10.13481/j.1671-587x.20150207

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

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. 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. 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/Abstract

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

燕蒙蒙, 徐珊, 高岩, 刘扬, 贺梦子, 陈司霖, 李鹏武, 刘晓冬, 马淑梅. miR-18a对结直肠癌SW116细胞辐射敏感性的影响[J]. 吉林大学学报(医学版), 2015, 41(02): 240-244.

YAN Mengmeng, XU Shan, GAO Yan, LIU Yang, HE Mengzi, CHEN Silin, LI Pengwu, LIU Xiaodong, MA Shumei. Effect of miR-18a on irradiation sensitivity of colorectal cancer SW116 cells[J]. Journal of Jilin University Medicine Edition, 2015, 41(02): 240-244.

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miR-18a对结直肠癌SW116细胞辐射敏感性的影响

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

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