RNAi沉默STAT3基因表达对Lewis肺癌细胞生长影响的实验研究  

Experimental study of the effect of RNAi to silence STAT3 gene expression on the growth of Lewis lung cancer cells

作　　者：王春光[1] 孙梅[2] 王荣有[1] 赵雪俭[3] 张兴义[1] 

机构地区：[1]吉林大学第二医院胸外科,吉林长春130041 [2]吉林大学第二医院病理科,吉林长春130041 [3]吉林大学基础医学院病理生理教研室

出　　处：《中国老年学杂志》2006年第2期210-213,共4页Chinese Journal of Gerontology

基　　金：吉林省科技厅资助课题(200505120)

摘　　要：目的探讨RNA干扰技术沉默转录信号传导子与激活子家族3(STAT3)对Lew is肺癌细胞凋亡、细胞生长抑制的影响。方法应用真核细胞转染技术将Psilencer2.1-U6-siRNA-STAT3重组质粒转入小鼠Lew is肺癌细胞,同时分别设立转染空质粒阴性对照组和转染试剂阴性对照。于72 h后收集细胞,分别提取细胞总蛋白及总RNA,分别用W estern b lot和RT-PCR测定STAT3基因在蛋白水平及mRNA水平的表达,用MMT法测定细胞的生长抑制状态,通过流式细胞仪检测Lew is肺癌细胞凋亡情况。结果重组质粒明显抑制STAT3蛋白的表达、降低STAT3 mRNA水平并诱导Lew is肺癌细胞凋亡、使肿瘤细胞生长明显受抑。结论Psilencer2.1-U6-siRNA-stat3 Lew is转染肺癌细胞后,可有效抑制STAT3蛋白表达和STAT3 mRNA的表达,诱导Lew is肺癌细胞凋亡和肿瘤细胞生长抑制。Objective Signal transducers and activators of transcription 3 （STAT3） silenced by RNA interference （RNAi） technique to induce the apoptosis and growth inhibition in Lewis lung cancer cells. Methods The recombinant plasmid Psilencer2.1-U6-siRNA-STAT3 was transfected into Lewis lung cancer cells by eukaryotic cell transfection technique. Plasmid vector and cell culture medium were used as negative and control group, respectively. The cells were collected 72 h later to extract cell total protein and RNA and to detect STAT3 gene protein and mRNA expression by Western blotting and RT-PCR. MTT method was used for the determinatian of cell growth inhibition. Apoptosis of Lewis lung cancer cell was determined with flowcytometry. Results The synthetic siRNA-STAT3 significantly inhibited STAT3 protein expression and decreased the levels of STAT3 mRNA, and induced apoptosis of Lewis lung cancer cells and inhibition of tumor growth. Conclusions The transfection of Lewis lung cancer cells by Psilencer2.1-U6-siRNA-STAT3 could effectively inhibit the expression of STAT3 protein and mRNA and induce apoptosis and growth inhibition in Lewis lung cancer cells.

关 键 词：RNA 干扰 STAT3 LEWIS肺癌 凋亡 

分 类 号：Q786[生物学—分子生物学]