J4 ›› 2010, Vol. 36 ›› Issue (5): 858-861.

• 基础研究 • 上一篇    下一篇

共表达siRNA-Stat3与GRIM-19质粒的构建及其功能研究

梁作文, 赵丽晶, 贾慧婕, 李馨, 高丽芳, 张灵, 赵雪俭   

  1. 吉林大学基础医学院病理生理学教研室 前列腺疾病防治研究中心|吉林 长春 130021
  • 收稿日期:2010-05-06 出版日期:2010-09-28 发布日期:2010-09-28
  • 通讯作者: 张 灵(Tel:0431-85619485,E-mail:zhangling3@jlu.edu.cn) E-mail:zhangling3@jlu.edu.cn
  • 作者简介:梁作文(1983-)|男|陕西省延安市人|在读医学博士|主要从事分子病理生理学研究。
  • 基金资助:

    国家自然科学基金资助课题(30973587);“十一五”重大科技专项基金资助课题 (2009ZX09304-003)

Construction of siRNA-Stat3 and GRIM-19 co-expression plasmid and study on its function

 LIANG Zuo-Wen, ZHAO Li-Jing, JIA Hui-Jie, LI Xin, GAO Li-Fang, ZHANG Ling, ZHAO Xue-Jian   

  1. Department of Pathophysiology,School of Basic Medical Sciences,Prostate Diseases Prevention and Treatment Research Center, Jilin University,Changchun 130021,China
  • Received:2010-05-06 Online:2010-09-28 Published:2010-09-28

摘要:

目的:构建共表达siRNA-Stat3和GRIM-19质粒,观察其对人前列腺癌PC-3M细胞增殖能力的影响。方法:以基因重组技术构建共表达siRNA-Stat3和GRIM-19质粒;应用真核细胞转染技术转染人前列腺癌PC-3M细胞,半定量RT-PCR法检测共表达质粒Stat3和GRIM-19 mRNA水平的变化,细胞增殖实验观察其对前列腺癌细胞增殖能力的影响。结果:酶切鉴定证实成功构建siRNA-Stat3和GRIM-19共表达质粒;与对照组比较,实验组前列腺癌细胞Stat3 mRNA表达显著降低,GRIM-19 mRNA表达显著升高(P<0.01);实验组前列腺癌细胞增殖率明显降低(P<0.05)。结论:成功构建siRNA-Stat3和GRIM-19共表达质粒,其对前列腺癌细胞株具有显著的生长抑制作用。

关键词: 前列腺肿瘤; Stat3;GRIM-19;基因治疗

Abstract:

Abstract:Objective
To construct recombinant plasmid co-expressing siRNA-Stat3 and GRIM-19,and observe its influence in the proliferation ability of prostate cancer cells.  Methods A recombinant plasmid co-expressing siRNA-Stat3 and GRIM-19 was constructed by using gene cloning. The prostate cancer cell lines PC-3M were transfected with the co-expression plasmid. The gene expression of Stat3 and GRIM19 was determined by semi-quantitative RT-PCR assay. The influence of recombinant plasmid in the proliferation ability of prostate cancer cells was determined by MTT. Results The siRNA-Stat3 and GRIM-19 co-expression plasmid was successfully constructed. Compared with control group,the expression of Stat3 mRNA was significantly decreased,and the expression of  GRIM-19 mRNA was significantly increased(P<0.01),the proliferation rate of prostate cancer cells was significantly decreased(P<0.05)in experimental group. Conclusion The siRNA-Stat3 and GRIM-19 co-expression plasmid  is successfully constructed,and  it has significant inhibition on the growth of prostate cancer cells.

Key words: prostate neoplasms;Stat3;GRIM-19;gene therapy

中图分类号: 

  • Q78