[Downregulation of MCL-1 by Diallyl Disulfide Induces G2/M Arrest in Human Leukemia K562 Cells and Its Mechanism].

Abstract

OBJECTIVE:

To investigate the inducing effect of down-regulation of MCL-1 by diallyl disulfide(DADS) on the G2/M arrest of human leukemia K562 cells and its mechanisms.

METHODS:

CCK-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.

RESULTS:

CCK-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 G2/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 G2/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).

CONCLUSION:

DADS can inhibit the K562 cell proliferation and induce them arrest G2/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.

• Authors: Xiao-Xia Ji, Fang Liu, Hong Xia, Jie He, Hui Tan, Lan Yi, Qi Su
• Journal: Zhongguo shi yan xue ye xue za zhi (Zhongguo Shi Yan Xue Ye Xue Za Zhi) Vol. 26 Issue 3 Pg. 750-755 (Jun 2018) ISSN: 1009-2137 [Print] China
• PMID: 29950215 (Publication Type: Journal Article)

Chemical References

• Allyl Compounds
• Disulfides
• Myeloid Cell Leukemia Sequence 1 Protein
• diallyl disulfide

Topics

• 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