Autophagy and apoptosis of acute myelogenous leukemia U937 cell induced by Sirolimus
10.3760/cma.j.issn.2095-428X.2015.17.014
- VernacularTitle:西罗莫司诱导急性髓系白血病U937细胞自噬及凋亡
- Author:
Wenfeng XU
;
Xiaoqin FENG
;
Chunfu LI
;
Xuedong WU
;
Yuelin HE
;
Yuming ZHANG
;
Fuyu PEI
- Publication Type:Journal Article
- Keywords:
Leukemia,myeloid;
U937 cells;
Sirolimus;
Autophagy;
Apoptosis
- From:
Chinese Journal of Applied Clinical Pediatrics
2015;30(17):1336-1340
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the autophagy and apoptosis in acute myelogenous leukemia U937 cell induced by Sirolimus.Methods U937 cells were subcultured, and blank control group(normal) and Sirolimus treated groups(12 h, 24 h,48 h) were established.The Sirolimus treated groups were treated by 2 μmol/L concentration of Sirolimus for 12 h,24 h and 48 h, respectively.The cell morphology of U937 cells treated by Sirolimus was observed after 12 h,24 h and 48 h.The survival rate of cells was detected by cell counting kit-8 method.Cell apoptosis was detected by flow cytometry using Annexin V-FITC/PI double labeled.Real-time PCR was used to detect the level of mRNA expression in autophagy specific protein maker mictotubule-associated protein light chain 3 (LC3)-Ⅱ in different treated times by Sirolimus.Sirolimus LC3 protein expression levels after treatment were detected by Western blot method.Results Under inverted microscope, the cell number of Sirolimus treatment group reduced gradually after 12 h ,24 h and 48 h culture, volume of cells became smaller, cells got ruptured, and the nucleus pycnosis and cellular debris increased.With the extension of time, U937 cells survival rate was falling, and there was statistical differences compared with those of the control group(P =0.031).With Sirolimus treatment, U937 cells after 12 h,24 h and 48 h, U937 cell apoptosis rate increased, and there were statistically significances, compared with those of the control group (P =0.027).With Sirolimus treatment U937 cells after 12 h,24 h and 48 h,LC3-Ⅱ mRNA expression and protein expression were down-regulated compared with those of the control group, and there were statistically significances (P =0.029).Conclusions Sirolimus can induce autophagy and apoptosis in U937 cells.Autophagy protein LC3-Ⅱ in gene and protein expression levels were lowered, and LC3-Ⅱ may play an important role in regulating the leukemia cell autophagy.
