The Study Of Anti-Human VEGF Ribozyme Treatment On Hepatocellular Carcinoma

Objective hepatocarcinoma cell line was transfected with anti-VEGF hairpin ribozyme gene mediated by Limpofectamine,To study the effect of anti-human VEGF hairpin ribozyme on the biological characteristic of hepatocarcinoma cell line SMMC-7721 and xenografted tumor of nude mice.Methods The ribozyme was synthesized and cloned into the eukaryotic express vector pcDNA3.1+ ,identified by double endonuclease digestion and DNA sequencing;After transfected into hepatocarcinoma cell by Limpofectamine,the RZ was screened by G418 and identified by RT-PCR and from genome level;MTT assay was carried to observe the effct of RZ on SMMC-7721 cell proliferation, The effect on VEGF expression of SMMC-7721cellline was conducted by semi-quantitative RT-PCR and ELISA;the cell apoptosis was analyzed by FCM. SMMC-7721/RZ and SMMC-7721/ pcDNA3.1+cell line was inoculated into nude rice,to see the changes of tumor volume and weight.MVD and VEGF expression was conducted by immunochemistry.Results The recombinant eukaryotic express vector pcDNA3.1+/RZ and the transgenic cell line SMMC-7721/RZ was successfully obtained,RZ can downregulate The level of VEGF expression, decline the growth rate of the hepatocarcinoma cell line and induce the cell apoptosis in a certain degree,VEGFexpression and angiogenesis of the xenografted tumor was obviously declined by the ribozyme.Conclusions anti VEGF hairpin ribozyme gene can inhibit the VEGF expression and...