Role of Pim1 Gene Overexpression in Pathogenesis of Acute Myeloid Leukemia.
10.19746/j.cnki.issn.1009-2137.2019.03.006
- Author:
Qing LI
1
;
Rui-Xue SUN
1
;
Yang OU
1
;
Hong-Mei LUO
1
;
Yu WU
2
Author Information
1. Department of Hematology, West China Hospital, Sichuan University, Laboratory of Hematology Research， Chengdu 610041, Sichuan Province, China.
2. Department of Hematology, West China Hospital, Sichuan University, Laboratory of Hematology Research， Chengdu 610041, Sichuan Province, China,E-mail: wu_yu@scu.edu.cn.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
Cell Proliferation;
Humans;
Leukemia, Myeloid, Acute;
genetics;
Proto-Oncogene Proteins c-pim-1;
genetics;
Signal Transduction;
U937 Cells
- From:
Journal of Experimental Hematology
2019;27(3):664-672
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effect of Pim1 expression up-regulation on cell proliferation, apoptosis, chemotaxis and angiogenesis in acute myeloid leukemia (AML) cell line U937, and to explore the possible molecular mechanism involved, finally to estimate the Pim1 expression in primary AML cells.
METHODS:GFP-tagged plasmid for Pim1 overexpression and an empty vector plasmid were constructed, and then a stable Pim1 expressed U937 cell line and a control virus-infected U937 cell line were established by a lentiviral vector system. After confirming Pim1 overexpression in U937 cells, proliferation and apoptosis are determined by CCK-8 Kit and flow cytometry respectively. Transwell chemotaxis assay was used to measure the effect of Pim1 overexpression on AML cells. Flow cytometry and confocal microscopy were applied to detect the influence of Pim1 overexpression on phosphorylated CXCR4 (pCXCR4) and its location. Real-time fluorescence quantitative PCR (qPCR) was used to detect the expression of angiogenesis and adhesion related genes in AML primary cells.
RESULTS:The lentivirus-infected AML cell line with Pim1 overex-pression and the control virusinfected AML cell line were established successfully. The Pim1 overexpression could enhance the proliferation and inhibit the cell apoptosis, moreover accompnied with the increasing expression of cyclin D1, phosphorylated BAD (pBAD) and pCXCR4. After SDF-1 α stimuli, Pim1 overexpression induced AML cell chemotaxis accompanied with p-CXCR4 expression and calcium influx increment. Pim1 overexpression has no effect on angiogenesis. Pim1 mRNA expression was significantly higher in AML patients than the healthy people.
CONCLUSION:Pim1 plays an important role in the pathogenesis of AML, which not only promotes AML cell proliferation and inhibition of apoptosis, but also enhances the chemotactic ability of leukemia cells, which closely relates with Pim1 phosphorylation of CXCR4 and the increase of intracellular calcium ion influx signals.
