RNA体外干扰肝癌Hep-3B细胞VEGF基因的表达

RNA interferes the expression of VEGF gene in hepatocarcinoma Hep-3B cells in vitro

目的:通过自主构建的RNAi载体和体外合成siRNA的方法,寻找能有效干扰VEGF基因表达的载体和siRNA片段治疗肿瘤.方法:将自主构建的pcDUVEGF-1和pcDUVEGF-2,以及体外合成的TWvegf-1和TWvegf-2 siRNA片段转染肝癌Hep-3B细胞,通过RT-PCR,Western Blot和ELISA检测转染前后VEGF的 mRNA及蛋白表达水平,以了解RNA干扰效果.结果:两种RNA干扰的方法均能抑制VEGF基因的表达且差异具有显著性( mRNA,0.28,0.47,0.76,0.58vs1.2,P<0.01;蛋白质,138,121,249,227vs389μg/L,P<0.01).结论:两个质粒及siRNA片段均能有效的抑制VEGF基因的表达.

AIM： To construct RNAi plasmids and synthesize siRNA in vitro, transport them into Hep-3B cells, so as to find some effective VEGF RNAi plasmids and siRNA segments. METHODS： The RNAi plasmids pcDUVEGF-1, pcDUVEGF-2 and siRNA segments TWvegf-1, TWvegf-2 were transported into Hep-3B cells, and the expressions of VEGF mRNA and protein in the transfected Hep-3B cells were checked using RT-PCR, Wes- tern Blot and ELISA. The results were compared between the transfected and the normal Hep-3B cells. RESULTS： There were prominent differences between the two groups in depressing the expression of mRNA （0.28, 0.47, 0.76, 0.58 vs 1.2, P 〈 0,01） and protein （138, 121, 249, 227 vs 389 i.~g/L, P 〈 0.01 ） of VEGF gene. CONCLUSION： The two RNAi plasmids and siRNA segments inhibit the expression of VEGF gene effec- tively,