Adenovirus-mediated siRNA inhibits the expression of VEGF-C in colorectal cancer cells

Objective To construct recombinant adenovirus containing small interfering RNA(siRNA) targeting human VEGF-C mRNA and observe the inhibitory effects of VEGF-C expression after transfecting LOVO cells.Methods The siRNA sequence targeting VEGF-C mRNA was synthesized and shuttle plasmid pDC316-VEGF-C siRNA-EGFP-U6 was constructed,which was co-transfected into 293 cells with the bone plasmid pBHGF35 to obtain the homologous recombination adenovirus Ad5F35-VEGF-C siRNA-EGFP-U6.The level of VEGF-CmRNA in the cells was tested by real time-PCR.The concentration of VECF-C protein in the cells was determined by Wetsern blot.Results The recombinant adenovirus Ad5F35-VEGF-C siRNA-EGFP-U6 was successfully constructed with the titre of 7.9×109IU/ml by purification.Ad5F35-VEGF-C siRNA-EGFP-U6 was transfected into LOVO cells.The expression of VEGF-C mRNA was decreased to the level of(30.7±1.2)% of the control(P0.05) at 48 hours after co-transfection,and the concentration of VEGF-C protein was decreased to the level of(47.8±2.2)% of the control at 72 hours after co-transfection(P0.05).Conclusion VEGF-C RNA interference mediated by adenovirus is capable of knocking down VEGF-C expression remarkablely in human colorectal cancer cells.