实用肝脏病杂志 ›› 2010, Vol. 13 ›› Issue (6): 401-403.doi: 10.3969/j.issn.1672-5069.2010.06.001

• 论著 •    下一篇

针对STAT3的siRNA抑制HepG2细胞增殖和p44/42MAPK蛋白的表达*

王新红,邢慧,陈晶,范玉晶,刘丹,刘红霞,吕志武   

  1. 150086 哈尔滨市 哈尔滨医科大学附属第二医院消化内科
  • 收稿日期:2010-07-21 出版日期:2010-12-10 发布日期:2016-04-18
  • 通讯作者: 吕志武,E-mail:drzwl@yahoo.com.cn
  • 作者简介:王新红 女,34岁,医学博士,主治医师。主要从事消化内科疾病的诊治及发病机制研究。E-mail:yuner1976@126.com
  • 基金资助:
    黑龙江省教育厅基金资助项目(编号:11531190);哈尔滨医科大学附属第二医院青年基金资助项目(编号:QN2007-17)

Inhibition of cell growth and p44/42MAPK protein expression by siRNA against STAT3 in HepG2 cells in vitro

WANG Xinhong,XING Hui,CHEN Jing,et al.   

  1. Department of Gastroenterology,Second Hospital Affiliated to Harbin Medical University,Harbin 150086,Heilongjiang Province,China
  • Received:2010-07-21 Online:2010-12-10 Published:2016-04-18

摘要: 目的 研究STAT3信号传导通路对HepG2细胞p44/42MAPK蛋白表达和细胞生长的影响。方法 将针对STAT3的siRNA转染入HepG2细胞以沉默STAT3基因的表达,采用MTT法检测细胞生长,采用Western blot法检测STAT3和p44/42MAPK蛋白的表达。结果 对照组和lipofectamineTM2000处理组细胞生长无明显差异(q=0.97,P>0.05),而siRNA处理组细胞生长被明显抑制(q=9.36,P<0.05);SiRNA转染后24h、48h、72h和96h,细胞抑制率分别为33.2%、39.6%、43.1%和33.9%,siRNA在96h后抑制作用减弱,细胞开始重新繁殖;转染细胞72h和96h后,可见STAT3蛋白表达均被抑制(t=14.12,P<0.05),p44/42MAPK蛋白表达未被抑制(F=3.99,P>0.05),而p-p44/42MAPK蛋白表达增加(t=16.30,P<0.05)。结论 STAT3信号传导通路可以影响细胞生长及p44/42MAPK蛋白质的磷酸化,而p-p44/42MAPK的表达增加可能代偿了沉默STAT3引起的细胞生长抑制,使细胞重新繁殖。

关键词: HepG2细胞, RNA干扰, 转录信号转导子与激活子3, P44/42丝裂素活化蛋白激酶, 信号传导通路

Abstract: Objective To study the effect of STAT3 signaling pathways on p44/42MAPK and cell growth in hepatocellular carcinoma. Methods The siRNA directed against STAT3 was transfected into HepG2 cells. Cell growth was measured by MTT assay and STAT3 and p44/42MAPK protein expression were measured by Western blot. Results A significant decrease in the number of viable cells(q=9.36,P<0.05)was found in the siRNA transfection group as compared to that in the control or the LipofectamineTM2000 transfected groups;SiRNA inhibited cell growth with the inhibition rates of 33.2% at 24h,39.6% at 48h,43.1% at 72h,and 33.9% at 96h;At 72h and 96h after the transfection,STAT3 protein expression was inhibited and P44/42MAPK protein expression was not influenced and p-p44/42MAPK protein expression was increased.Conclusion siRNA against STAT3 may affect cell growth in vitro.

Key words: HepG2 cell, RNA interference, Signal transducers and activators of transcription 3, p44/42MAPK, Signaling pathway