Effects of inhibition of Cripto gene siRNA on vascular endothelial growth factor of colon cancer cell line LS-174T
- VernacularTitle:RNAi沉默Cripto基因对结肠癌细胞血管内皮生长因子的抑制
- Author:
Yu FAN
;
Youli ZHANG
;
Hua LI
;
Zefeng XU
;
Shu ZHENG
- Publication Type:Journal Article
- Keywords:
Colonic neoplasms;
Genes,Cripto;
RNA interence;
siRNA;
Vascular endothelial growth factors
- From:
Chinese Journal of Pathophysiology
1986;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To study the effects of Cripto gene on vascular endothelial growth factor(VEGF) of colon carcinoma cells.METHODS: Cripto siRNA was designed and constructed.Colon cancer LS-174T cells were divided into 4 groups: control group and different dose (3.125,6.25 and 12.5 nmol/L) of siRNA groups.After transfected for 24,48 and 72 h,colon cancer cells were harvested to carry on the next tests.Expression of Cripto mRNA was determined with real-time PCR,and immunofluorescence isothiocyanate(FITC) labeling assay and Northern blotting were performed to examine the expression of protein and mRNA of VEGF,respectively.The cells in control group and cells transfected with 12.5 nmol/L siRNA were inoculated into nude mice respectively.30 days after inoculated,the mice of two groups were executed,and immunohistochemical(ICH) assay was used to evaluate the VEGF protein of mice tumor.RESULTS: siRNA down-regulated the Cripto mRNA in a dose and time dependent manner.Protein and mRNA of VEGF in transfected cells reduced in a dose and time dependent manner.Compared to control,the expression of VEGF protein from ICH assay was lowered significantly(P
