Objective: We propose to block the autocrine and (or) paracrine pathway of vascular endothelial growth factor (VEGF) in ovarian cancer by using anti-VEGF hairpin ribozyme gene to see whether the growth of tumor cells could be inhibited and to further exploit its mechanisms.
Methods: Anti-VEGF hairpin ribozyme gene eukaryotic expression vector was introduced into ovarian cancer cells SKOV(3) by using lipofectin mediation and positive clones were screened by genticin (G418); ribozyme (RZ) expression was confirmed by RNA dot blot; the VEGF expression of SKOV(3) cells before or after transfection were detected by reverse transcription polymerase chain reaction (RT-PCR) method, transmission electron microscopy demonstrated the morphologic changes of ribozyme-generating SKOV(3) cells; the growth of transfected cells in nude mice were detected.
Results: VEGF expression was decreased distinctly in SKOV(3) cells transfected by RZ (SKOV(3)-RZ) cells. Apoptosis cells were observed; the formation rate and growth speed of xenografted tumor slowed down.
Conclusions: Anti-VEGF hairpin ribozyme gene can inhibit the expression of VEGF mRNA and VEGF in SKOV(3) cells. The growth of xenografted tumor in nude mice was inhibited by reduced angiogenesis. This provides an experimental basis for curing human ovarian cancer with antiangiogenesis method.