Effect of VEGF-C interfering on the lymphangiogenesis of a mouse model bearing human colon cancer / 肿瘤研究与临床

ABSTRACT

Objective To construct recombinant adenovirns containing small interfering RNA (siRNA) targeting human VEGF-C mRNA,then to study the inhibitory effects of adenovirus-mediated VEGF-C siRNA on growth and lymphangiogenesis of human colon cancer in BABL/c nude mice model.Methods In vitrothe specific siRNA sequence targeting human VEGF-C mRNA was selected.The homologous double-strand DNA was designed and synthesized.After such DNA was inserted into pDC316-EGFP-U6 by BamHI and HindⅢ,pDC316-VEGF-C siRNA-EGFP-U6 was obtained,then it was co-transfected into 293 cells with the hone plasmid pBHGF35.After generated by homologous recombination,Ad5F35-VEGF-C siRNA-EGFP-U6 was obtained,and it was packaged and amplified in 293 cells.The human colon cancer model was established in nude mice by hypo-injection LoVo cells.They were divided randomly into four groups (n=6)adenovirus,virus without target gene,single siRNA and PBS control group.Injecting intervention intra-tumorly in each groups,calculating the tumors volume and drawing the growth curve,calculating micro-vascular density (MVD) and micro-lymphatic density (LVD) by staining respectively of the lymphatic and vascular with anti-LYVE-1 monoclonal antibody and anti-CD<,34> monoclonal antibody were completed.Results Adenovirus group tumor sizes were smaller than other groups.The LVD were 8.47±2.1 and 17.35±4.7 (P<0.05) in adenovirns group and the PBS control group.The MVD were 22.65±6.04 and 23.19±7.63 (P>0.05) in adenovims group and the PBS control group respectively.Conclusion The adenovirus-mediated VEGF-C siRNA can significantly inhibit the growth and lymphangiogenesis of human colon cancer in nude mice model,hut have no effect on microangium vessels growth.