Livin基因和Survivin基因联合靶向siRNA诱导结肠癌细胞凋亡的作用

Apoptosis induction effect of siRNA recombinant expression vector targeting Livin and Survivin gene simultaneously on human colon cancer cells

目的探讨Livin基因和Survivin基因联合靶向小干扰（si）RNA对结肠癌细胞增殖及凋亡的影响。方法构建Livin和Survivin联合靶向的siRNA重组表达载体并转染结肠癌细胞．通过RT—PCR和蛋白质印迹方法分别检测Livin和Survivin的表达．通过MTT法检测siRNA对细胞增殖的抑制作用．利用流式细胞仪检测处理后细胞的凋亡效应。结果经酶切鉴定和测序分析证实Livin和Survivin联合靶向的siRNA重组表达载体构建成功。这种联合靶向SiRNA对Livin mRNA及蛋白表达的抑制率分别为27．9％和22．3％．对Survivin mRNA及蛋白表达的抑制率分别为32．2％和40．9％。与对照组相比，联合靶向siRNA可降低肿瘤细胞增殖率．增加细胞凋亡率，但其细胞生长抑制作用和促细胞凋亡作用均弱于单独干扰Livin或Survivin基因．结论Livin和Survivin基因联合靶向siRNA能降低结肠癌细胞中Livin和survivin基因的表达．抑制结肠癌细胞的增殖．并诱导结肠癌细胞的凋亡．但此协同抑制作用较单独干扰Livin或survivin基因为弱。

Objective To investigate the effect of siRNA targeting Livin and Survivin gene simuhaneously on the proliferation and apoptosis of human colon cancer cells. Methods SiRNA recombinant expression vectors targeting Livin and Survivin gene simultaneously were constructed and transfected into human colon cancer cell line Lovo. The effects of siRNA recombinant expression vector on Lovo cells were detected by RT-PCR, Western blot, MTT reduction assay and flow cytometry. Results It was confirmed by restriction endonuclease and sequence analysis that siRNA recombinant expression vector targeting Livin and Survivin gene simultaneously was constructed successfully. The suppressive rates of siRNA targeting Livin and Survivin gene simultaneously on Livin mRNA and protein expression were 27.9% and 22.3% respectively, and those on Survivin mRNA and protein expression were 32.2% and 40.9% respectively. The survival rate of cancer cells was decreased whereas the apoptotic rate was increased, but the coordinate repression was weaker than Livin and Survivin RNA interference alone. Conclusions siRNA targeting Livin and Survivin gene simultaneously can decrease the expression of Livin and Survivin gene, suppress cell proliferation and induce cell apoptosis in human colon cancer. The coordinate repression was weaker than Livin and Survivin RNA interference alone.