Li Guo-Qing, Zhang Yu, Shen Wei-Gan, Zhou Wei, Gao Na, Gu Jian
Department of Hematology and Rheumatolagy, Clinic medical College, Yangzhou University, Yangzhou 225001, Jiangsu Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2008 Feb;16(1):38-43.
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.
本研究旨在探讨金雀异黄素(gen)对氯化钴(CoCl₂)诱导的人白血病细胞系K562中缺氧诱导因子-1α(HIF-1α)表达的影响。采用CoCl₂模拟缺氧条件;制备剂量效应组和时间效应组如下:剂量效应组分别用0、50、100和150 μmol/L的CoCl₂处理72小时,时间效应组用100 μmol/L的CoCl₂处理,分别在0、24、48和72小时进行检测。将经gen处理的样本分为五组:(1)正常对照组;(2)150 μmol/L CoCl₂组;(3)150 μmol/L CoCl₂ + 50 μmol/L gen组;(4)150 μmol/L CoCl₂ + 100 μmol/L gen组;(5)150 μmol/L CoCl₂ + 200 μmol/L gen组。分别采用RT-PCR和Western blot检测HIF-1α mRNA和蛋白。结果表明,CoCl₂诱导的K562细胞中HIF-1α蛋白表达呈剂量和时间依赖性增加(p<0.01),而K562细胞中HIF-1α mRNA表达水平保持不变(p>0.05)。Gen能以剂量依赖性方式显著抑制CoCl₂诱导的HIF-1α蛋白表达(p<0.01),而gen处理对HIF-1α mRNA表达无影响(p>0.05)。结论:CoCl₂剂量和时间依赖性诱导HIF-1α蛋白表达;无论常氧条件还是常氧条件下存在钴离子,HIF-1α mRNA均持续表达。Gen可在蛋白水平抑制CoCl₂诱导的K562细胞中HIF-1α表达,但对mRNA无影响。