摘要
目的探讨缺氧时肝癌细胞SMMC-7721中乏氧诱导因子-1α(HIF-1α)、己糖激酶II(HKII)表达及对细胞周期调控机制的影响。方法人肝癌细胞SMMC-7721分成3组:缺氧12h组、缺氧24h组及对照组。缺氧组分别在缺氧条件下(37℃,5%CO2、2%O2饱和度)培养12h和24h,对照组置于正常氧浓度条件下(37℃,5%CO2、21%O2饱和度)培养24h。流式细胞仪检测细胞周期分布,免疫组织化学SP法检测HIF-1α表达,荧光定量PCR法检测HKIImRNA表达量。结果缺氧状态下细胞周期分析处于G0/G1期细胞百分比明显增多,处于G2/M期的细胞减少,并且可见到较明显的凋亡峰的形成,缺氧12h组、缺氧24h组的G0/G1期比例显著高于对照组(P<0.05);HIF-1α和HKII在两组缺氧组与对照组比较,其表达量明显增加(P<0.05),缺氧24h组与缺氧12h组比较差异有统计学意义(P<0.05)。结论缺氧导致细胞阻滞在G0/G1期,HIF-1α可刺激人肝癌细胞HKII表达的增加,以HIF-1α为药物靶点可望成为治疗肝癌的重要新方法。
Objective To investigate the impact of cell cycles arrest, expression of hypoxia-inducible factor-1α （ HIF-1α ） and hexokinaseII（HKII） in hepatocellular carcinoma cells under the hypoxic environment. Methods Human hepatocellular carcinoma cell line SMMC-7721 were divided into 3 groups： group of hypoxia for 12h and for 24h, and the control group. The two hypoxic groups were exposed to hypoxic condition（37℃, 5% CO2, 2% O2） for 12h and 24h respectively. The control group was exposed to normal oxygen condition（37℃, 5% CO2, 21% O2） for 24h. Cell cycle was examined by flow cytometry analysis; HIF-1α was measured by immuno- histochemistry and HKII was detected by real-time quantitative PCR. Results Cells accumulated in the gap 0 and gap 1 （G0/G1） phases （groups of hypoxia for 24h and 12h vs the control group, P〈O.05）; while reduced in the gap2 and mitosis （G2/M） phases of the cell cycle, and the apoptosis peak significantly occurred after hypoxia treatment. HIF-1α and HKII expression was significantly higher in hypoxic groups than that in control group （groups of hypoxia for 24h and 12h vs the control group, P〈0.05; group of hypoxia for 24h vs for 12h, P〈0.05）. Conclusion A marked G0/G1 arrest of cell cycle and a high level of HKII expression are induced under the hypoxic environment, demonstrating the potential role of HIF-1α -targeted therapy in hepatocellular carcinoma.
出处
《中华普通外科学文献（电子版）》
2008年第2期20-22,共3页
Chinese Archives of General Surgery(Electronic Edition)
关键词
缺氧
缺氧诱导因子-Α
己糖激酶
肝癌
Hypoxia
Hypoxia-inducible factor-1α
Hexokinase II
Hepatocellular carcinoma
