Suppression of vascular endothelial growth factor expression by adenovirus-mediated RNA interference in treatment of lung adenocarcinoma cells / 第二军医大学学报

ABSTRACT

Objective: To investigate the inhibitory effects of RNA silencing via adenovirus mediated vascular endothelial growth factor(VEGF) shRNA on proliferation of lung adenocarcinoma cells in vitro and in vivo. Methods: PCR method was used to construct a pAd Easy/VEGF adenovirus vector containing enhanced green fluorescent protein (EGFP) gene and expressing VEGF shRNA. The 293 cells were transfected with the linearized pAd Easy/VEGF using Lipofectamine2000. Then lung adenocarcinoma cells A459 were transfected with the constructed vector. The EGFP expression was detected by fluorescent microscopy and flow cytometry. VEGF mRNA expression was examined by RT-PCR and Western blotting. The cell growth was observed with MTT method and the growth curve was plotted. Meanwhile, nude mice were transplanted with A549 cells to establish tumor models and the growth of tumors were observed. Results: The recombinant pAd-Easy carrying shRNA targeting VEGF had been constructed and the aim sequence had been obtained. The transfection efficiencies in pAd-Easy/VEGF and blank vector transfected A549 cells were 100% and 99.7%, respectively. RT PCR and Western blotting showed a remarkable decrease of VEGF expression in the pAd-Easy/VEGF group compared with normal saline group. The tumor growth in pAd-Easy/VEGF group was obviously slowed down and the weight and volume of tumors were both significantly lower than those of the control group (all P<0.01). Conclusion: The shRNA targeting VEGF constructed in the present study can efficiently decrease the VEGF expression in A549 cells in vitro and suppress the growth of A549 cells in vivo.