Effects of RNA interference on PIDD expression and drug resistance of HT-29 cells
- VernacularTitle:RNA干扰对HT29细胞PIDD蛋白表达及细胞耐药性影响的研究
- Author:
Jingjing PENG
;
Jianjun LI
;
Houjie LIANG
- Publication Type:Journal Article
- Keywords:
colonic neoplasms;
PIDD protein;
HT29 cells;
RNA interference;
drug resistance,neoplasms
- From:
Medical Journal of Chinese People's Liberation Army
2001;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the distributive changes of PIDD protein in HT29 cells stimulated by 5-fluorouracil(5-FU),and the influence of the PIDD protein interfered by small interference RNA(siRNA) on the drug resistance of HT29 cells in vitro.Methods siRNA was used to interfere the PIDD expression,and HT-29 cells were treated with 5-FU.Western blotting was employed to detect the PIDD expression before and after interference.The distribution of PIDD in nucleus and cytoplasm after 5-FU treatment was also detected.Changes in drug sensitivity of HT-29 cells to 5-FU were determined with MTT assay and IC50 values were evaluated.Results PIDD protein was expressed mainly in the cytoplasm before 5-FU stimulation,and it migraded into the nucleus after stimulation.Western blotting showed that the total PIDD expression and the expression both in cytoplasm and nuclei decreased significantly after RNA interference,and no increase of the total PIDD and that migrated into nucleus was found even after 5-FU stimulation.It was found that in the transfection group(cells were incubated 12h after being transfected with PIDD-360),normal group(cells without treatment) and control group(cells were incubated 12h after being transfected with negative transfection reagent),the IC50 values of cells treated with 5-FU were 0.23?0.06?g/ml,2.71?0.70?g/ml and 2.78?1.03?g/ml,respectively.The IC50 value declined significantly in transfection group compared with that of both the normal group and control group(P
