Downregulation of MCL-1 by Diallyl Disulfide Induces G/M Arrest in Human Leukemia K562 Cells and Its Mechanism.
- Author:
Xiao-Xia JI
1
,
2
;
Fang LIU
3
;
Hong XIA
3
;
Jie HE
3
;
Hui TAN
3
;
Lan YI
3
;
Qi SU
4
Author Information
- Publication Type:Journal Article
- MeSH: Allyl Compounds; Apoptosis; Cell Line, Tumor; Disulfides; Down-Regulation; G2 Phase Cell Cycle Checkpoints; Humans; K562 Cells; Leukemia; M Phase Cell Cycle Checkpoints; Myeloid Cell Leukemia Sequence 1 Protein
- From: Journal of Experimental Hematology 2018;26(3):750-755
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the inducing effect of down-regulation of MCL-1 by diallyl disulfide(DADS) on the G/M arrest of human leukemia K562 cells and its mechanisms.
METHODSCCK-8 was used to detect the effect of DADS on proliferation of K562 cells, flow cytometry was employed to observe the effect of cycle arrest by DADS and RNAi silencing MCL-1 gene in K562 cells. The expressions of MCL-1, PCNA and CDK1 in K562 cells treated with DADS were detected by Western blot. The amphigamy of MCL-1 with PCNA and CDK1 was detected by Coimmunoprecipitation.
RESULTSCCK-8 detection showed that the inhibition rates of K562 cells treated with 15, 30, 60, 120, 240 µmol/L DADS were 32.48%, 59.34%, 66.42%, 77.06%, 81.05% respectively (P<0.05). Flow cytometry analysis revealed that the perecentages of G/M cells were increased to 18.6% and 34.4%, 17.5% and 28.5%, respectively at 24 and 48 h after treating K562 cells with 60 and 120 µmol/L DADS (P<0.05). And the perecentage of G/M cells of silencing MCL-1 was significantly increased (P<0.05). Silencing effects of MCL-1+DADS on the cells were enhanced more significantly as compared with DADS or MCL-1 alone (P<0.05). Western blot exhibited that DADS could markedly downregulate the expression of MCL-1, PCNA and CDK1(P<0.05). Coimmunoprecipitation revealed that MCL-1 bound with PCNA and CDK1, then forming heterodimers, which were downregulated respectively more significantly than that in the control group after treating K562 cells with DADS for 8 h (P<0.05).
CONCLUSIONDADS can inhibit the K562 cell proliferation and induce them arrest G/M through downregulation of MCL-1, then decreasing the expression of PCNA and CDK1 in hunan leukemia K562 cells. Moreover, silencing MCL-1 can enhance the effect of DADS.