靶向生存素基因的小干扰RNA重组表达载体诱导人结肠癌细胞凋亡

Induction of apoptosis of human colon cancer cells by siRNA recombinant expression vector targeting survivin gene.

作者信息

Cai Ming, Wang Guobin, Tao Kaixiong, Cai Changxue

机构信息

Department of General Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.

出版信息

J Huazhong Univ Sci Technolog Med Sci. 2009 Feb;29(1):45-9. doi: 10.1007/s11596-009-0109-1. Epub 2009 Feb 18.

DOI:10.1007/s11596-009-0109-1

PMID:19224161

Abstract

This study constructed siRNA recombinant expression vector targeting survivin gene and observe the apoptosis induction effect of it in human colon cancer cells. siRNA recombinant expression vector targeting survivin gene was constructed and transfected into human colon cancer cells. The effect of siRNA recombinant expression vector was detected by RT-PCR, Western blot, MTT reduction assay and flow cytometry. It was confirmed by restriction endonuclease and sequence analysis that siRNA recombinant expression vector targeting survivin gene was constructed successfully. Inhibition rate of survivin siRNA at mRNA and protein levels was 36.33% and 44.65% respectively. Growth of cancer cells was inhibited and the apoptosis rate was (17.24+/-2.13)%. The siRNA recombinant expression vector targeting survivin gene has been constructed successfully. It not only can inhibit the expression of survivin gene, but also can induce apoptosis in human colon cancer cells remarkably.

摘要

本研究构建了靶向survivin基因的siRNA重组表达载体，并观察其对人结肠癌细胞的凋亡诱导作用。构建了靶向survivin基因的siRNA重组表达载体并转染到人结肠癌细胞中。通过RT-PCR、Western blot、MTT比色法和流式细胞术检测siRNA重组表达载体的作用。经限制性内切酶和序列分析证实成功构建了靶向survivin基因的siRNA重组表达载体。survivin siRNA在mRNA和蛋白水平的抑制率分别为36.33%和44.65%。癌细胞生长受到抑制，凋亡率为(17.24±2.13)%。成功构建了靶向survivin基因的siRNA重组表达载体。它不仅能抑制survivin基因的表达，还能显著诱导人结肠癌细胞凋亡。