Effects of hypoxia-inducible factor inhibitor on expression of HIF-1alpha and VEGF and induction of apoptosis in leukemic cell lines.
- Author:
Fei WANG
1
;
Bao-An CHEN
;
Jian CHENG
;
Wen-Lin XU
;
Xue-Mei WANG
;
Jia-Hua DING
;
Chong GAO
;
Yun-Yu SUN
;
Jun WANG
;
Gang ZHAO
;
Wen BAO
;
Hui-Hui SONG
;
Feng GAO
;
Wei ZHANG
;
Guo-Hua XIA
;
Xiao-Ping PEI
;
Wei-Wei WU
;
Li YIN
;
Xue-Yun SHAN
Author Information
1. Department of Hematology, Zhongda Hospital, Southeast University Clinical Medical College, Nanjing 210009, Jiangsu Province, China.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
drug effects;
Cell Hypoxia;
Gene Expression Regulation, Leukemic;
Humans;
Hypoxia-Inducible Factor 1, alpha Subunit;
antagonists & inhibitors;
metabolism;
Indazoles;
pharmacology;
Jurkat Cells;
K562 Cells;
U937 Cells;
Vascular Endothelial Growth Factor A;
metabolism
- From:
Journal of Experimental Hematology
2010;18(1):74-78
- CountryChina
- Language:Chinese
-
Abstract:
This study was purposed to investigate the effect of a hypoxia-inducible factor inhibitor (YC-1) on expression of hypoxia-inducible factor 1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF) as well as induction of apoptosis in leukemic cell lines. RT-PCR was used to determine the levels of HIF-1alpha mRNA and VEGF mRNA in K562, U937 and Jurkat cells. After treatment of U937 cell with 4 micromol/L YC-1, cell apoptosis was assayed by DAPI staining under fluorescent microscope and flow cytometry with Annexin V-FITC/PI staining; the expression levels of HIF-1alpha mRNA and VEGF mRNA were measured with RT-PCR; the expression levels of HIF-1alpha, VEGF, BAX, BCL-2 and caspase-3 proteins were measured by Western blot. The results showed that HIF-1alpha mRNA and VEGF mRNA were expressed in all three leukemia cell lines. After treatment of U937 cell with 4 micromol/L YC-1 for 0, 8, 16 and 24 hours, the changes of morphologic features of U937 cells could be observed under fluorescent microscope and the apoptotic rates significantly increased in time-dependent manner, they were (4.87 +/- 0.70)%, (27.27 +/- 2.00)%, (51.53 +/- 2.81) and (60.5 +/- 3.20)% respectively, the expression levels of VEGF mRNA reduced, while the expression levels of HIF-1alpha mRNA had no obviously changes.Furthermore, the expression of HIF-1alpha, VEGF and BCL-2 decreased, while the expression of BAX and caspase-3 increased, the ratio of BAX/BCL-2 increased in time-dependent manner (r = 0.973, p < 0.01). It is concluded that HIF-1alpha mRNA and VEGF mRNA are all expressed in in K562, U937 and Jurkat cells, YC-1 has significant effect on down-regulating the protein expression of HIF-1alpha and VEGF, and induces the apoptosis in U937. The mechanism of apoptosis in leukemic cells may involve in up-regulating BAX/BCL-2 ratio and expression of protein caspase-3.
