Induction of Apoptosis and Upregulation of Fas Ligand and Fas - Associated Death Domain by Doxorubicin in Human Jurkat Leukemia T- Cells
- VernacularTitle:阿霉素诱导Jurkat白血病细胞凋亡及上调Fas配体和Fas相关死亡域
- Author:
Jia-hao, LIU
;
Hong-li, TANG
;
Wei-yong, RUAN
;
Wei, WANG
;
Li, HUANG
- Publication Type:Journal Article
- Keywords:
leukemia;
apoptosis;
flow cytometry;
Western blot;
FasL;
Fas associated death domain;
caspase
- From:Journal of Applied Clinical Pediatrics
2006;21(1):53-56
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the dose and time kinetics of induction of apoptosis induced by doxorubicin in J urkat leukemiacells, and to explore its pertinent molecular mechanisms. Methods Human Jurkat leukemia T - cells were treated with doxorubicin at theconcentration of 0.1 mg/L, 0.2 mg/L, 0.5 mg/L and 1.0 mg/L for 6,12,24 and 36 hours, respectively, of which one sample was pre-treated with zVAD- fmk (benzyloxycarbonyl - Val -Ala - Asp - fluoromethylketone) prior to addition of doxorubicin 0.2 mg/L. Apop-tosis was detected with both annexin V - FITC and propidium iodide ( PI ) staining and annexin V FITC and PI double positive cellswere analyzed by flow cytometry. Western blot was used to evaluate the level of Fas ligand (FasL) and FADD (Fas - associated death do-main) expression. Results The differences of apoptotic cells induced by all dose of doxorubicin were not significant (P＞0.05 ) at 6hour;at 12 hour, only the highest dose, 1 mg/L, significantly induced cell apoptosis;while the lowest dose,0.1 mg/L, did not significantlycaused cell apoptosis for all time points. After exposure to the doses of 0.2 and 0.5 mg/L for 24 or 36 hours,a significant increase in per-centage of apoptotic cells was observed (P ＜ 0.001 ). Apoptosis induced by doxorubicin was completely inhibited when the cells were incu-bated with doxorubicin in the presence of zVAD - fmk (P ＜ 0. 001 ). The level of FasL and FADD expression correspondingly increasedwith exposure time to doxorubicin. Conclusions Doxorubicin induces apoptosis in a dose - and time - dependent manner; upregulatedFasL may initiate the activation of the Fas signaling pathway and caspases are the ultimate executioner in the induction of leukemia cellapoptosis by doxorubicin.
