摘要
目的 探讨应用RNA干扰(RNAi)技术沉默信号转导子与转录活化子(STAT3)基因对膀胱癌细胞株T24及5637细胞的影响.方法 针对STAT3 mRNA序列设计合成3对编码小干扰RNA(siRNA)的DNA模板,构建pGenesil-1-shRNA-STAT3重组质粒,转染人类膀胱癌细胞株T24及5637细胞.通过半定量RT-PCR、Western blotting法检测STAT3基因不同水平的表达情况,采用流式细胞仪检测转染后细胞周期的变化.结果 成功构建pGenesil-1-shRNA-STAT3重组质粒,并成功转染T24及5637膀胱癌细胞;半定量RT-PCR、Western blotting结果显示,重组质粒转染组细胞的STAT3基因表达在RNA及蛋白水平上显著低于对照组(P<0.05);流式细胞仪的结果显示,重组质粒转染组细胞明显受到抑制并出现凋亡现象.结论 pGeneSil-1-shRNK-STAT3转染膀胱癌细胞株T24及5637细胞后,可有效抑制STAT3的表达,并抑制膀胱癌细胞的生长.
Objective Signal transducers and activators of transcription 3 (STAT3) silenced by RNA interference (RNAi) technique were used to induce the apoptosis and growth inhibition in T24 and 5637 bladder cancer cells. Methods Three recombinant plasmids pGenesil-1-shRNA-STAT3 was constructed and transfected into T24 and 5637 cells. The expression of STAT3 gene was detected by RT-PCR and Western blotting. FCM was used to observe the apoptosis in T24 and 5637 cells. Results pGenesil-1-shRNA-STAT3 was successfully constructed, and transfected into T24 and 5637 cells. RT-PCR and Western blot analysis demonstrated that pGenesil-1-shRNA-STAT3 could significantly inhibit the expression of STAT3 in T24 and 5637 cells; FCM results show that it could suppress the growth of 1'24 and 5637 cells. Conclusion pGeneSiI-1-shRNA-STAT3 could significantly inhibit STAT3 expression, suppress the growth of T24 and 5637 cells.
出处
《肿瘤研究与临床》
CAS
2008年第6期372-375,共4页
Cancer Research and Clinic
