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World J Gastroenterol. Feb 21, 2006; 12(7): 1140-1143
Published online Feb 21, 2006. doi: 10.3748/wjg.v12.i7.1140
Inhibitory effect of antisense vascular endothelial growth factor RNA on the profile of hepatocellular carcinoma cell line in vitro and in vivo
Ji-Hui Hao, Ming Yu, Hui-Kai Li, Yu-Rong Shi, Qiang Li, Xi-Shan Hao
Ji-Hui Hao, Hui-Kai Li, Yu-Rong Shi, Qiang Li, Xi-Shan Hao, Department of Abdominal Surgery, Tianjin Cancer Hospital, Tianjin 300060, China
Ming Yu, Public Health College, Tianjin Medical University, Tianjin 300070, China
Co-first authors: Ming Yu
Supported by the Natural Science Foundation of Tianjin, No 013615611
Correspondence to: Dr. Ji-Hui Hao, Department of Abdominal Surgery, Tianjin Cancer Hospital, Hexi District, Tianjin 300060, China. jihuihao@yahoo.com
Telephone: +86-22-23340123-3325
Received: June 13, 2005
Revised: July 2, 2005
Accepted: July 12, 2005
Published online: February 21, 2006
Abstract

AIM: To evaluate the effect of antisense vascular endothelial growth factor (VEGF) RNA (PCMV-FGEV) transfection on the profile of hepatocellular carcinoma (HCC) SMMC-7721 cells in vitro and in vivo.

METHODS: SMMC-7721 cells were transfected with PCMV-FGEV antisense, PCMV-VEGF sense and empty vector plasmid encapsulated by lipofectamine as antisense group, sense group and control group respectively. The positive cell clones were selected with G418. The stable transfection and expression of VEGF in the cells were determined by RT-PCR and immunohistochemistry. Cell proliferation was observed by MTT assay. FACS analysis was used to determine the effect of PCMV-FGEV transfection on cell apoptosis. The growth of transfected cells in vivo was also observed in nude mice.

RESULTS: VEGF expression was reduced in SMMC-7721 transfected with PCMV-FGEV, which was confirmed by RT-PCR and immunohistochemistry. No effect of PCMV-FGEV transfection was found on cell proliferation and cell apoptosis of SMMC-7721 in vitro. The growth of cells transfected with PCMV-FGEV was slow in nude mice and accompanied with obvious apoptosis. The latent time of tumors in the antisense group was 25.0 ± 1.8 d, which was longer than that in sense and control groups (F = 19.455, P < 0.01). The average tumor weight in antisense group (0.96 g ± 0.28 g) was the smallest among the three groups (F = 21.501, P < 0.01).

CONCLUSION: The expression of VEGF can be inhibited by antisense PCMV-FGEV. Antisense PCMV-FGEV has no effect on cell proliferation and apoptosis of SMMC-7721 in vitro but can inhibit tumor growth and induce cell apoptosis in vivo.

Keywords: Antisense RNA, Vascular endothelial growth factor, Gene expression, Hepatocellular carcinoma, Transfection