Experimental study on shRNA targeted hTERT gene to suppress bladder cancer cell growth
- VernacularTitle:shRNA下调人端粒酶逆转录酶基因抑制膀胱癌细胞生长作用的实验研究
- Author:
Penghui ZHANG
;
Lin ZOU
;
Zhiguang TU
- Publication Type:Journal Article
- Keywords:
Bladder neoplasms;
Carcinoma;
hTERT
- From:
Chinese Journal of Urology
2001;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the mechanism of siRNA targeted hTERT gene to inhibit bladder cancer T24 cell growth by decreasing c-myc and TGF-?1 expression. Methods shRNA-hTERT-pTZU6+1 vectors were constructed by RNAi-DNA vector technique, then the vectors were transfected into bladder cancer T24 cells,and the most effective vector and its optimal concentration were screened using RT-PCR to detect hTERT expression in T24 cells.The T24 cell growth, the alternative of cell phase,the expression of hTERT,c-myc and TGF-?1 were detected by flow cytometry,RT-PCR and immunohistochemistry. Results Three shRNA-hTERT-pTZU6+1 vectors were successfully constructed.The most effective vector was ph2-shRNA vector,and its optimal concentration was 1.0 ?g.This vector decreased the cell growth and the cell number of S phase from 65.2% to 38.6%,increased the cell number of G0/G1 phase from 32.0% to 57.9%,and attenuated both mRNA and protein expressions of hTERT,c-myc and TGF-?1 in T24 cells. Conclusions Targeted hTERT gene with siRNA may inhibit the cell proliferation of bladder cancer;down-regulating hTERT expression by attenuating the expression of c-myc and TGF-?1 is probably involved in the mechanism.
