Effects of oxygen and calcium on the expression of wild-type and mutated hypoxia-inducible factor-1alpha eukaryotic vectors in HEK293 cells.
- Author:
Rui-Bin FU
1
;
Ping-Sheng WU
;
Jian QIU
;
Ding-Cheng XIANG
;
Jun MA
;
Chang-Jiang HONG
;
Jian-Xin HE
Author Information
- Publication Type:Journal Article
- MeSH: Calcium; metabolism; Cell Hypoxia; Genetic Vectors; HEK293 Cells; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; genetics; metabolism; Oxygen; metabolism; RNA, Messenger; genetics; Transfection; Vascular Endothelial Growth Factor A; metabolism
- From: Journal of Southern Medical University 2009;29(7):1321-1323
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the effects of oxygen and calcium on the expression of eukaryotic vectors harboring wild-type or mutated hypoxia-inducible factor-1alpha (HIF-11alpha) in HEK293 cells.
METHODSHEK293 cells were transiently transfected with pcDNA3.1+/HIF-11alpha, pcDNA3.1+/HIF-11alpha-564Ala and pcDNA3.1+/HIF-11alpha-564Ala-803Ala via lipofectin. Western blotting were used to detect HIF-11alpha protein after normoxic or hypoxic exposure of the transfected HEK293 cells in the presence or absence of Ca(2+). The levels of vascular endothelial growth factor (VEGF) mRNA in the transfected cells in normoxic condition was detected using RT-PCR.
RESULTSThe levels of HIF-11alpha protein and VEGF mRNA increased in HEK293 cells transfected with the vectors harboring mutated HIF-11alpha, but not in the cells transfected with wild-type HIF-11alpha vectors in normoxia. Hypoxia increased the levels of HIF-11alpha protein in the cells transfected with wild-type HIF-11alpha vectors, which was inhibited by the application of Ca(2+). Ca(2+) showed no inhibitory effect on HIF-11alpha levels in HEK293 cells transfected with the vectors containing mutated HIF-11alpha.
CONCLUSIONThe protein products of pcDNA3.1+/HIF-11alpha-564Ala and pcDNA3.1+/HIF-11alpha- 564Ala-803Ala in HEK293 cells enhance the cell tolerance to oxygen and protease.
