摘要
目的:观察氯化钴(CoCl2)模拟缺氧环境下肝癌细胞生物钟基因表达的变化。方法:以不同浓度(0,50,100,200μmol/L)的CoCl2处理肝癌细胞系Huh7细胞24 h后,用Western blot检测细胞中缺氧诱导因子1α(HIF-1α)的蛋白表达,real-time PCR检测细胞中生物钟基因CLOCK、BMAL1、Per1、Per2、Per3、Cry1、Cry2、CKIε的mRNA表达。结果:Western blot结果显示,未处理Huh7细胞(0μmol/L CoCl2)无HIF-1α表达,而各浓度的CoCl2处理后,Huh7细胞出现明显的HIF-1α的蛋白表达,且表达水平随CoCl2浓度增加而升高。real-time PCR结果显示,CoCl2处理后的Huh7细胞中BMAL1、Per1、Cry2的mRNA水平明显上调,而CLOCK、Cry1、Per2、Per3、CKIε的mRNA水平明显下调,且均呈CoCl2浓度依赖性(均P<0.05)。结论:缺氧微环境可能是引起肝癌细胞生物钟基因表达异常的原因之一。
Objective： To observe the expression changes of the clock genes in hepatocellular carcinoma （HCC） cells under cobalt chloride （CoCl2）-induced hypoxic condition. Methods： HCC Huh7 cells were exposed to different concentrations （0, 50, 100 and 200 txrnol/L） of CoC12 for 24 h, and then, the protein expressions of hypoxia-inducible factor-let （HIF-la） in the cells were determined by Western blot analysis and the mRNA expressions of the clock genes that included CLOCK, BMAL1, Perl, Per2,Per3, Cry1, Cry2 and CKIε in the cells were detected by real-time PCR method. Results： The results of Western blot showed that there was no visible HIF-la protein expression in the Huh7 cells （0 μmol/L CoCl2）, while the HIF-lα protein expressions were clearly seen in the Huh cells treated with each concentration of CoClv and the expression level was increased with CoCl2 concentration,The results of real-time PCR showed that the mRNA levels of BMAL 1, Per1 and Cry2 were up-regulated while mRNA levels of CLOCK, Cry1, Per2 and CKIε mRNA levels were down-regulated significantly after CoCl2 treatment, and all presented a CoC12 concentration-dependent manner （all P〈0.05）. Conclusion： Hypoxia microenvironment may be one of the causes for the abnormal expression of clock genes in HCC cells.
出处
《中国普通外科杂志》
CAS
CSCD
北大核心
2014年第1期48-52,共5页
China Journal of General Surgery
关键词
癌
肝细胞
缺氧
肝癌
生物钟
缺氧诱导因子1
Α亚基
Carcinoma, Hepatocellular
Anoxia
Biological Clocks
Hypoxia-Inducible Factor i, alpha Subunit
