c-Myc 靶向小分子干扰RNA 抑制人直肠癌Colo320 细胞增殖的实验研究

中国临床药理学与治疗学 ›› 2006, Vol. 11 ›› Issue (11): 1301-1304.

c-Myc 靶向小分子干扰RNA 抑制人直肠癌Colo320 细胞增殖的实验研究

黄浩, 凃欣¹, 余南才, 刘倩, 易艳东, 马威

武汉第一医院中心实验室, 武汉 430022, 湖北;
¹华中科技大学人类基因组研究中心, 武汉 430074, 湖北

收稿日期:2006-06-02 修回日期:2006-06-29 出版日期:2006-11-26 发布日期:2020-11-06
作者简介:黄浩, 男, 博士, 主治医师, 从事病原生物学与肿瘤分子生物学研究。Tel:027-85855908-455 E-mail:whhuanghao10@yahoo.com.cn
基金资助:
湖北省技攻关计划项目( No2006AA301B66-3)

Use siRNA targeting gene c-Myc to inhibit cellular proliferation in human rectal cancer Colo320 cell line

HUANG Hao, TU Xin¹, YU Nan-cai, LIU Qian, YI Yang-dong, MA Wei

Center of Experimental Medicine, Wuhan First Hospital , Wuhan 430022 , Hubei , China;
¹Human Genome Research Center, Huazhong University of Science &Technology , Wuhan 430074 , Hubei, China;

Received:2006-06-02 Revised:2006-06-29 Online:2006-11-26 Published:2020-11-06

摘要/Abstract

摘要： 目的利用小分子干扰RNA( siRNA) 技术抑制人直肠癌Colo320 细胞c-Myc 基因的表达。为研究c-Myc 基因在人直肠癌Colo320 细胞中的作用提供一个新的方法。方法设计人直肠癌Colo320 细胞基因特异性小分子干扰RNA, 用体外转录方法合成人直肠癌Colo320 细胞基因的小分子干扰RNA 并转染人直肠癌Colo320 细胞, 培养48 ~ 96 h 后, 收集细胞RNA 应用实时荧光定量RT-PCR 方法检测转染细胞中c-Myc 基因mRNA 水平变化, Western blot 检测C-MYC 蛋白的表达, 四甲基偶氮唑蓝( MTT 法) 和集落形成实验检测细胞增殖活性。结果转染siRNA后, 与对照组相比, 实验组pGensil-c-Myc-1-4 的c-Myc 基因mRNA 水平明显降低, MTT 法和集落形成实验表明实验组的增殖速率明显低于对照组的增殖速率。结论在人直肠癌Colo320 细胞中存在RNA干扰的机制, 特异性siRNA 能够有效的抑制c-Myc基因的表达, 为研究c-Myc 基因在肿瘤细胞中的调节途径提供了一个新的方法。

关键词: 小分子干扰RNA, 人直肠癌, c-Myc 基因

Abstract: AIM: To inhibit c-Myc gene expression in human rectal cancer cell line Colo320 by small interfering RNA ( siRNA) , and to provide a new method to study the role of c-Myc in Colo320 cell.METHODS: The c-Myc gene specific siRNA was designed according to its sequence and made by in vitro transcription.The c-Myc siRNA was transfected into Colo320 cell, and the cell was cultured for 48 to 96 hours before harvesting.c-Myc mRNA and protein level were monitored by using fluorescence real time reverse transcription polymerase chain reaction.Cellular proliferation activities were assayed by tetrazolium bromide (MTT) colorimetry and clone test.RESULTS: Compared with control group, there was a significant decrease in the c-Myc mRNA.Colo320 cells transfected with c-Myc siRNA had lower cellular proliferation than no-transfected Colo320 cells.CONCLUSION: Our results suggest that there exist RNA interference mechanism in Colo320 cell line, and the c-Myc siRNA can specifically inhibit the expression of c-Myc gene in Colo320 cells.RNA interference method provides a new way to study the role of c-Myc in cancer cell.

Key words: small interfering RNA, human rectal cancer, c-Myc gene

中图分类号:

R965.2
R966

黄浩, 凃欣, 余南才, 刘倩, 易艳东, 马威. c-Myc 靶向小分子干扰RNA 抑制人直肠癌Colo320 细胞增殖的实验研究[J]. 中国临床药理学与治疗学, 2006, 11(11): 1301-1304.

HUANG Hao, TU Xin, YU Nan-cai, LIU Qian, YI Yang-dong, MA Wei. Use siRNA targeting gene c-Myc to inhibit cellular proliferation in human rectal cancer Colo320 cell line[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2006, 11(11): 1301-1304.