Construction of antisense VEGF eukaryotic expression vector and effect to ovarian cancer cell / 中国癌症杂志

ABSTRACT

Purpose:To constract and identify eukaryotic expression vector carrying human VEGF cDNA to cure human ovarian cancer with antiangiogenesis method. Methods:Reverse transcription PCR for VEGF, VEGF was inserted into eukaryotic exprssion vector pcDNA3 to construct pcDNA3-VEGF(-), in which restriction enzyme analysis was used to confirm the reverse orientation fo the VEGF cDNA from the individual transformants. The vector was transfected into human ovarian cancer cell HO-8910 and the positive clone was screened by G418,VEGF expression was confirmed by RNA dot blot. The VEGF expression of HO-8910 cells before or after transfection was detected by Western blot and imunofluorescence.The biological characteristics of HO-8910 cells before and after transfectionwas inspected. Results: VEGF gene was obtained by RT-PCR, the pcDNA3-VEGF(-) vector was obtained. VEGF expression was blocked by anisense VEGF RNA. The clone formation ability of singal cell was decreased after transfected, G 1 phase cells were increased and S Phase celIs were decreased in cell cycle. The proliferation of HO-89l0 cell was reduced. The formation rate and growth speed of xengrafted tumor slowed down. Conclusions:The successful construction of antisense VEGF eukaryotic expression vector is of significance for ovarian cancer-specific antisense gene therapy.