Effect of antisense VEGF 1⁶⁵cDNA transfection on the growth of human neuroblastoma SH-SY5Y cells in vitro and in nude mice

Objective: The present study was designed to explore the inhibitory effect of antisense VEGF₁₆₅ cDNA on angiogenesis, growth rate of human neuroblastoma. Methods: The eukaryotic expression vectors bearing antisense VEGF₁₆₅ cDNA or sense VEGF₁₆₅ cDNA were constructed. The stable cell lines transfected with the sense or antisense VEGF₁₆₅ cDNA were established by using the selective medium containing 400 mg/L of G418. These cell lines were further studied for the exogenous antisense VEGF mRNA expression by RT-PCR and inhibition of expression of endogenous VEGF protein by immunocytochemical. staining and ELISA. The proliferation of the transfected cells was analyzed by MTT method. Transfected cells were subcutaneously transplanted into nude mice and the growth of tumor masses was observed and weighted. Results: The antisense VEGF was only detected in SH-SY5Y/AsVEGF cells by RT-PCR. The expression of VEGF protein was dramatically declined in the SH-SY5Y/AsVEGF cells compared with the original cells and empty vector transfected cells by immunocytochemical staining and ELISA. There was no effect of antisense VEGF transfection on cell proliferation in vitro. The growth of tumor mass with the cells transfected with antisense VEGF was significantly decreased in nude mice. Conclusion: An tisense VEGF cDNA transfection to SH-SY5Y cells can effectively inhibit the expression of endogenous VEGF protein and tumor growth in nude mice.