Study on inhibitory effect of antisense VEGF RNA on the growth of hepatocellular in vitro in vivo.
- Author:
Ji-hui HAO
1
;
Ming YU
;
Yu-rong SHI
;
Qiang LI
;
Xi-shan HAO
Author Information
- Publication Type:Journal Article
- MeSH: Apoptosis; Cell Division; Cell Line, Tumor; Humans; Immunohistochemistry; Liver Neoplasms; pathology; therapy; RNA, Antisense; therapeutic use; Transfection; Vascular Endothelial Growth Factor A; analysis; antagonists & inhibitors; genetics
- From: Chinese Journal of Hepatology 2003;11(12):725-727
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo determine the effects of PCMV-FGEV transfection on the profile of SMMC-7721 hepatocellular in vitro in vivo.
METHODSSMMC-7721 hepatocellular was transfected with PCMV-FGEV antisense, PCMV-VEGF sense and empty vector plasmid encapsulated by lipofectamine. The positive cell clones were selected with G418. The stable transfection and expression of VEGF in the SMMC-7721 hepatocellular were determined by in situ hybridization and immunochemical analysis. The effect of PCMV-FGEV transfection on SMMC-7721 hepatocellular proliferation was observed by MTT colorimetric assay. Flow cytometry was used to determine the effects of PCMV-FGEV transfection on cell apoptosis of SMMC-7721. The growth of transfected cells was also observed in nude mice.
RESULTSThere was reduced VEGF expression in SMMC-7721 transfected with PCMV-FGEV confirmed by in situ hybridization and immunohistochemical analysis. There was no effect of PCMV-FGEV transfection on cell proliferation and cell apoptosis of SMMC-7721 in vitro. The growth of cell with PCMV-FGEV transfected was slow in nude mice (vivo) and accompanied with obvious apoptosis. The latent time of tumor in the antisense mice group was 25.0+/-1.8 days, which was longer than that in sense and control group significantly (F=19.455, P<0.01). On the other hand, 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).
CONCLUSIONSThe expression of VEGF was inhibited by PCMV-FGEV. There was no effect on cell proliferation and cell apoptosis of SMMC-7721 by transferring PCMV-FGEV gene into SMMC-7721 cells in vitro. But in vivo it can inhibit tumor growth and induce cell apoptosis.