[Inhibitory effect of genistein on hypoxia-inducible factor-1alpha expression induced by cobalt chloride in leukemia cell line K562].
Guoqing Li, Yu Zhang, Weigan Shen, Wei Zhou, Na Gao, Jian Gu
Abstract
Guoqing Li, Yu Zhang, Weigan Shen, Wei Zhou, Na Gao, Jian Gu
Abstract
This study was aimed to investigate the effect of genistein (gen) on the expression of hypoxia inducible factor-1alpha (HIF-1alpha) induced by cobalt chloride (CoCl(2)) in human leukemia cell line K562. The hypoxia condition was simulated by CoCl(2); the dose- and time-effect groups were prepared as follows: the former were exposed to 0, 50, 100 and 150 micromol/L of CoCl(2) for 72 hours, the latter were detected at 0, 24, 48 and 72 hours while treated with CoCl(2) 100 micromol/L. The gen-treated samples were divided into five groups: (1) normal control; (2) CoCl(2) 150 micromol/L; (3) CoCl(2) 150 micromol/L + gen 50 micromol/L; (4) CoCl(2) 150 micromol/L + gen 100 micromol/L; (5) CoCl(2) 150 micromol/L + gen 200 micromol/L. The HIF-1alpha mRNA and protein were detected by RT-PCR and Western blot respectively. The results indicated that the expression of HIF-1alpha protein in K562 cells induced by CoCl(2) increased in dose-and time-dependent manner (p<0.01), while the expression of HIF-1alpha mRNA in K562 cell remained the similar level (p>0.05). Gen significantly inhibited the expression of HIF-1alpha protein induced by CoCl(2) in dose-dependent manner (p<0.01) while the HIF-1alpha mRNA expression was not affected by treatment of gen (p>0.05). It is concluded that CoCl(2) dose- and time-dependently induced the HIF-1alpha protein expression; HIF-1alpha mRNA was constantly expressed regardless of normoxic conditions or in the presence of cobalt ion under normoxic conditions. Gen can inhibit HIF-1alpha expression in K562 cell induced by CoCl(2) at level of protein, but not mRNA.
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This study was aimed to investigate the effect of genistein (gen) on the expression of hypoxia inducible factor-1alpha (HIF-1alpha) induced by cobalt chloride (CoCl(2)) in human leukemia cell line K562. The hypoxia condition was simulated by CoCl(2); the dose- and time-effect groups were prepared as follows: the former were exposed to 0, 50, 100 and 150 micromol/L of CoCl(2) for 72 hours, the latter were detected at 0, 24, 48 and 72 hours while treated with CoCl(2) 100 micromol/L. The gen-treated samples were divided into five groups: (1) normal control; (2) CoCl(2) 150 micromol/L; (3) CoCl(2) 150 micromol/L + gen 50 micromol/L; (4) CoCl(2) 150 micromol/L + gen 100 micromol/L; (5) CoCl(2) 150 micromol/L + gen 200 micromol/L. The HIF-1alpha mRNA and protein were detected by RT-PCR and Western blot respectively. The results indicated that the expression of HIF-1alpha protein in K562 cells induced by CoCl(2) increased in dose-and time-dependent manner (p<0.01), while the expression of HIF-1alpha mRNA in K562 cell remained the similar level (p>0.05). Gen significantly inhibited the expression of HIF-1alpha protein induced by CoCl(2) in dose-dependent manner (p<0.01) while the HIF-1alpha mRNA expression was not affected by treatment of gen (p>0.05). It is concluded that CoCl(2) dose- and time-dependently induced the HIF-1alpha protein expression; HIF-1alpha mRNA was constantly expressed regardless of normoxic conditions or in the presence of cobalt ion under normoxic conditions. Gen can inhibit HIF-1alpha expression in K562 cell induced by CoCl(2) at level of protein, but not mRNA.
Key concepts: Cobalt chloride, K562 cells, Chemistry, Western blot, Molecular biology, Messenger RNA, Cell culture, Hypoxia (environmental)