山东医药2018，Vol.57Issue(7)：5-8,4.DOI:10.3969/j.issn.1002-266X.2018.07.002
缺氧诱导因子抑制剂YC-1对缺氧肝癌细胞生物钟基因和蛋白表达及增殖的影响
Effects of hypoxia-inducible factor inhibitor YC-1 on expression and proliferation of circadian clock genes and proteins in hypoxic hepatoma cells
摘要
Abstract
Objective To observe the effects of hypoxia inducible factor(HIF)inhibitor YC-1 on the circadian clock genes,circadian clock proteins and cell proliferation of hepatocellular carcinoma(HCC)cells.Methods The HCC Huh7 cells were cultured in the hypoxia incubator and then were divided into the experimental group and control group. Cells in the the experimental group were added with 50 μmol/L YC-1,and cells in the control group were added with DM-SO.At 48 h after culture,the mRNA and protein of two groups were exacted,respectively.The real-time PCR was used to detect the Per1,Per2,Per3,Cry1,Cry2,Clock, and Bmal1 mRNA, while the Per1, Per2, Per3,Cry1, Cry2, Clock, and Bmal1 proteins were detected by Western blotting in cells of the two groups.The Huh7 cells in the logarithmic growth phase were divided into the experimental group and control group.YC-1(50 μmol/L)was added into the experimental group,while the same amount of 0.9%NaCl solution was added into the control group.Cells were cultured under hypoxic conditions.Clone formation experiment was used to observe the proliferation,and then the clone formation rate was calcu-lated.Results The relative mRNA and protein expression levels of Clock, Per1, Per3, and Cry1 in the experimental group were lower than those in the control group,but the relative mRNA and protein expression levels of Bmal 1,Per2,and Cry2 were higher than those in the control group(all P<0.05).The number of clones was 83 and the colony formation rate was 27.67%in the experimental group, versus 191 and 63.67% in the control group, respectively.The number of colony formation and colony formation rate in the experimental group were all lower than those in the control group(all P<0.05).Conclusions After YC-1 treatment,the expression of Clock,Per1,Per3,and Cry1 genes and proteins is down-regulated in the HCC cells,while the expression of Bmal1,Per2,and Cry2 genes is up-regulated,and the cell proliferation and colony forming abilities decrease.关键词
缺氧诱导因子抑制剂YC-1/肝细胞癌/生物钟/生物钟基因/细胞增殖Key words
hypoxia/hypoxia-inducible factor inhibitor YC-1/hepatocellular carcinoma/circadian clock/circadian clock gene/cell proliferation分类
医药卫生引用本文复制引用
许静,孙诚谊,喻超,何晓晓,杨盛力..缺氧诱导因子抑制剂YC-1对缺氧肝癌细胞生物钟基因和蛋白表达及增殖的影响[J].山东医药,2018,57(7):5-8,4.基金项目
国家自然科学基金青年基金项目(81702885) （81702885）
湖北省自然科学基金项目(2017CFB178) （2017CFB178）
中央高校基本科研业务费专项资金项目:华中科技大学自主创新基金项目(0118530328) （0118530328）
中国肝炎防治基金会天晴肝病研究基金项目(TQGB20170005). （TQGB20170005）
