摘要
目的观察小干扰RNA（siRNA）联合介导Livin和Survivin基因沉默对人肾癌细胞786-0增殖抑制及凋亡的作用。方法采用实时荧光定量聚合酶链反应（FQ-PCR）及Western blot分别检测肾癌细胞株和组织中Livin和Survivin的表达。构建Livin和Suvivin联合靶向的siRNA重组表达载体短发卡RNA（shRNA），转染后检测786-0中Livin和Suvivin的表达变化。待出现明显干扰效果后采用细胞计数试剂盒（CCK-8）和流式细胞仪分别检测重组表达载体shRNA对786-0的增殖抑制及凋亡诱导影响。结果联合介导组较分别单独介导组比较增殖抑制效应增强23％、26％；凋亡诱导增强100％、116％。结论siRNA联合介导Livin和Survivin基因表达下调，发挥协同增强的siRNA作用，表现为更强的增殖抑制和凋亡诱导作用。
Objective To investigate the effect of combine mediated Livin and Survivin genes silencing induced the proliferation inhibition and apoptosis in human renal carcinoma cell line 786-0. Methods Application of real-time fluorescent quantitative polymerase chain reaction （FQ-PCR） and Western blotting methods respectively, to detect the expression level of Livin and Sarvivin in the renal cancer cell lines and tissues. Constructed Livin and Survivin combine targeting small interfering RNA （siRNA） recombinant expression vector, then transfected it into 786-0 cell line and validation. Application of the cell counting kit-8 （CCK-8） test and flow cytometer to investigate the proliferation inhibition and apoptosis of 786-0. Results The cell survival rate of the group of combine mediated Livin and Survivin genes silencing was significant decrease and then the cell apoptotie rate was significant increase, the proliferation inhibition enhanced 23%, 26% and apoptosis increased 100%, 116%, respectively. Conclusion siRNA combine mediated Livin and Survivin gene expression, give nlav to the role of synergistic enhancement of siRNA.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2014年第6期1199-1201,共3页
Chinese Journal of Experimental Surgery
