The Effect of si-PKM2 on Proliferation and Apoptosis of Acute Leukemic Cells and Its Molecular Mechanism.
10.19746/j.cnki.issn.1009-2137.2021.05.005
- Author:
Li-Yuan LI
1
;
Zi-Yuan NIE
1
;
Xiao-Yan ZHANG
1
;
Jian-Min LUO
2
;
Lin YANG
1
;
Qian WANG
1
;
Xing-Zhe WANG
1
Author Information
1. Department of Hematology, The Second Hospital of Hebei Medical University, Hebei key Laboratory of Hematology, Shijiazhuang 050000, Hebei Province, China.
2. Department of Hematology, The Second Hospital of Hebei Medical University, Hebei key Laboratory of Hematology, Shijiazhuang 050000, Hebei Province, China E-mail: ljm315@163.com.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
Cell Proliferation;
Glycolysis;
Humans;
Phosphatidylinositol 3-Kinases/metabolism*;
Pyruvate Kinase
- From:
Journal of Experimental Hematology
2021;29(5):1394-1402
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effect of glycolytic enzyme pyruvate kinase type 2 (PKM2) on the proliferation and apoptosis of human leukemia HL-60 cells.
METHODS:si-PKM2 plasmid was transfected into HL-60 cells (set as si-PKM2 group), and blank vector transfected cells were set as control group (si-Ctl group). The expression levels of PKM2 mRNA and protein in si-Ctl group and si-PKM2 group were detected by RT-qPCR and Western blot. CCK-8 cell detection kit was used to detect the proliferation ability of the cells in the two groups. Flow cytometry was used to detect the changes of cell cycle and apoptosis. Western blot and RT-qPCR were used to detect the changes of p-Akt and p-mTOR protein levels in PI3K/Akt/mTOR signaling pathway and the changes of glycolysis-related mRNA levels of the cells in the two groups. The changes in glucose consumption and lactic acid production of the cells were assayed. Over expressed PKM2, HL-60 cells were treated with PI3K inhibitor LY294002 or galactose, the changes in cell proliferation ability, cell cycle and apoptosis, as well as changes in glucose consumption and lactic acid production were detected.
RESULTS:Interfered by si-PKM2, mRNA and protein levels of PKM2 in si-PKM2 group significantly decreased, and proliferation ability of the cells was also reduced (P<0.05). After PKM2 knockdown, the cells were significantly blocked at G
CONCLUSION:PKM2 knockdown can inhibit the proliferation and induce apoptosis of HL-60 cells, and its molecular mechanism may be related to the PKM2-mediated PI3K/Akt/mTOR-glycolysis, which suggesting that PKM2 may serve as a molecular target for the prevention and treatment of leukemia.
