Effects of PIM1 Gene on Proliferation,Apoptosis and JAK2/STAT3 Signaling Pathway of Acute Myeloid Leukemia U937 Cells
Objective:To investigate the effects of the serine/threonine kinase family member 1(PIM1)gene on the proliferation and apoptosis of acute myeloid leukemia(AML)U937 cells,and the regulation effect on Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)pathway.Methods:Bone marrow mononuclear cells from newly diagnosed adult AML patients and patients with iron deficiency anemia were collected and PIM1 mRNA expression was detected by RT-qPCR.AML cell line U937 cells were divided into U937 group(U937 cells were cultured normally),Si-PIM1 group(U937 cells were transfected with low expression adenovirus vector containing PIM1 mRNA),Si-NC group(U937 cells were transfected with low expression adenovirus vector without PIM1 mRNA),coumermycin A1(CoA1)group(JAK2 activator CoA1 was added to U937 cells at a concentration of 20 μ mol/L),and Si-PIM1+CoA1 group(U937 cells were transfected with adenoviral vector containing low expression of PIM1 mRNA and added with CoA1 at a concentration of 20 μ mol/L).After culture for 24 h,the expressions of PIM1 mRNA and protein,JAK2/STAT3 pathway,cell cycle and apoptosis-related proteins in U937 cells were detected by RT-qPCR and Western blot,the cell proliferation activity was detected by MTT assay,and flow cytometry was used to detect cell cycle changes and apoptosis rate.Results:The PIM1 mRNA expression level in bone marrow mononuclear cells in AML patients was higher than that in patients with iron deficiency anemia(P＜0.05).Compared with U937 group,PIM1 mRNA and protein,phosphorylated JAK2(p-JAK2)/JAK2,phosphorylated STAT3(p-STAT3)/STAT3,Cyclin D1,cyclin-dependent kinase 2(CDK2)protein,cell proliferation activity,S phase and G2/M phase proportions were decreased in Si-PIM1 group(all P＜0.05),while p27,Caspase-3 protein,G0/G,phase proportion and apoptosis rate were increased(all P＜0.05).However,the changes of above indicators in CoA1 group were just opposite to those in Si-PIM1 group,indicating that CoA1 could reverse the effect of Si-PIM1 on U937 cells.There were no significant differences in above indexes of U937 cells between U937 group,Si-PIM1+CoA1 group and Si-NC group(P＞0.05).Conclusion:Knockdown of PIM1 gene expression can inhibit U937 cell proliferation and promote apoptosis,in order to alleviate ALM process,which may be related to the inhibition of JAK2/STAT3 pathway activation.
serine/threonine kinase family member 1acute myeloid leukemia U937 cellsproliferationapoptosisJanus kinase 2/signal transducer and activator of transcription 3 pathway
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