Abstract:

BACKGROUND: Previous studies demonstrated that proliferation of cancer cells can be inhibited via RNA interference on the expression of vascular endothelial growth factor (VEGF). However, few studies report RNA interference on the expression of VEGF in gallbladder carcinoma.
OBJECTIVE: To design and screen shRNA targeting VEGF, and to observe the effect of small interfering RNA targeting on proliferation of gallbladder cancer cells.
METHODS: The VEGF-shRNA fragment was synthetized and connected with pCYU6/GFP/Neo-shRNA plasmid vector. shRNA was transfected into gallbladder cancer cells. The gallbladder carcinoma models of nude mice were prepared and randomly divided into blank control, negative control and experimental groups, with 6 animals in each group. shRNA was injected into tumor. Cell growth was detected by fluorescence microscope MTT. The RNA interference efficiency was examined by fluorescent quantitative RT-PCR. Changes of tumor volume were also observed.
RESULTS AND CONCLUSION: Gallbladder cancer cells were shrunk with round shapes and a part of cells were dead after RNA interference on VEGF. shRNA-VEGF1 and shRNA-VEGF2 could significantly inhibit mRNA gene expression of VEGF, the inhibition ratio was 86% and 82%, respectively. The tumor volume of the experimental group was smaller than the other groups, with slowly growth (P < 0.05). No obvious changes were found in the blank control and negative control groups. The constructed hVEGF-shRNA vector markedly decreases VEGF gene expression and inhibits cellular proliferation, eventually, to treat gallbladder cancer.