Role of receptor-interaction proteins in phenethylisothiocyanate-induced apoptosis in human leukemia cells in vitro
- VernacularTitle:受体作用蛋白在异硫氰酸苯乙酯诱导人急性白血病细胞凋亡中的作用
- Author:
Jie YAO
;
Ning GAO
- Publication Type:Journal Article
- Keywords:
phenethylisothiocyanate;
leukemia;
apoptosis;
receptor-interaction proteins
- From:Journal of Third Military Medical University
1983;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the role of receptor-interaction proteins(RIP) in phenethylisothiocyanate(PEITC)-induced apoptosis in human leukemia cells.Methods U937 cells were exposed to PEITC at various concentrations of 0,2,4,6 and 8 ?mol/L for 3 and 6 h,or at 8 ?mol/L for different time intervals of 1,3,6,9,12 and 24 h.Cells were stained with Annexin V/PI,and apoptosis was determined by using flow cytometry.Total protein extracts were prepared and subjected to Western blot assay using antibodies against poly(ADP-ribose)polymerase(PARP),Caspase-3,Caspase-8,and RIP.?-actin was used to ensure equivalent loading.Results After exposure to PEITC at concentrations of 2,4,6 and 8 ?mol/L for 3 h,U937 cells showed an increase in cell apoptosis(9% ,25% ,44% and 56% respectively).While when the time expanded to 6 h,the amount of apoptotic cells was increased to 16% ,28% ,51% and 82% respectively.A dose-dependent manner was seen when U937 cells were exposure to PEITC at different concentrations.When U937 cells were treated by PEITC at concentration of 8 ?mol/L for 1,3,6,9,12,and 24 h,the apoptotic cells covered 12% ,57% ,79% ,86% ,90% ,and 91% of all cells respectively,indicating a time-dependent manner.Western blot assay showed that PEITC caused the activation/cleavage of Caspase-3 and-8,and improved the PARP cleavage,and also decreased the expression of RIP.PEITC-induced apoptosis was proceeded by degradation of RIP.Conclusion Our results indicate that PEITC induces the apoptosis in human leukemia cells through a process that involves death receptor pathway,and RIP may play an important role in the apoptosis.
