摘要
选择血管内皮生长因子(VEGF)基因为靶基因,设计两组针对VEGFmRNA的小干扰RNA,合成DNA寡核苷酸链,体外转录合成siRNA.以人大肠癌细胞系HT-29为靶细胞,应用脂质体转染的方法,将siRNA导入细胞.MTT法检测siRNA对细胞增殖率的影响,RT-PCR法比较转染前后VEGFmRNA表达水平的变化,ELISA法检测细胞培养液中VEGF蛋白分泌量的变化.结果表明,两组siRNA转染后均能有效地抑制HT-29细胞的生长,VEGFmRNA的表达量大幅度减少;相对应的VEGF蛋白水平也显著降低,而作为阴性对照的错义序列组siRNA转染后则无上述作用.
Designed specific small interfering RNA（siRNA） targeting VEGF mRNA and synthesized oligo fragments, then small interfering RNA were obtained by in vitro transcription and transfected into cultured human colon carcinoma cell line HT-29 with lipofectin. Analyzed the effect of the siRNA on proliferation of HT-29 cells by MTT assay and expression level of VEGF mRNA of transfected cells by RT-PCR as well as amount of secreted VEGF protein in the supernatant by ELISA. Two groups of siRNA targeting human VEGF effectively inhibited proliferation of HT-29 cells after transfected; secretion of VEGF protein also notably decreased,but the control scramble siRNA showed no effect.
出处
《生命科学研究》
CAS
CSCD
2005年第4期356-360,共5页
Life Science Research
基金
青岛市科技局科技计划项目(03-1-YN-14)
