Effect Of Vascular Endothelial Growth Factor Antisense RNA On Myeloma Cell U266 Growth And Its Angiogenesis

Objective:To investigate the feasibility of antisense RNA technique targeting at VEGF gene in the treatment of multiple myeloma.Methods:VEGF121 cDNA was inserted into multiple clone site of eukaryotic expression vector pIRES2-EGFP to construct AS-VEGF. Then AS-VEGF or pIRES2-EGFP vector control was transferred into human myeloma cells U266 with lipofectamine 2000. U266 cells stably transfected with AS-VEGF or pIRES2-EGFP were acquired by drug selection with G418.The expression of VEGF mRNA and protein in U266-AS or U266-V were asssayed by RT-PCR or western blotting .The viability and apoptosis of U266-AS or U266-V cells in serum-free RPMI1640 were observed by MTT assay and flow cytometry(FCM),respectively.The proliferation and migration of vascular endothelial cells ECV304 in the present of the conditioned medium (CM) from U266-AS or U266-V cell were observe by MTT assay and transwell chamber method,respectively.Results:The eukaryotic expression vector AS-VEGF expressing VEGF antisense RNA were constructed successfully.Expression of VEGF antisense RNA in U266 cells reduced the production and secretion of VEGF protein ,inhibited U266 survival in serum-free RPMI1640.The proliferation and migration of vascular endothelium cell ECV304 were inhibited significantly in the present of CM from U266-AS cells than that from U266-V cells.Conclusions:These results suggest that VEGF antisense RNA could inhibit the production of VEGF of myeloma cells , perform anti-myeloma and anti-angiogenesis in vitro. VEGF antisense RNA therapy may constitute a novel treatment strategy of multiple myeloma.