【摘要】 目的探讨甲胎蛋白(AFP)对正常肝细胞和肝癌细胞生物钟基因表达的影响。方法将AFP过表达的慢病毒载体感染正常肝细胞L02细胞和肝癌细胞株SMMC-7721细胞构建AFP过表达的正常肝细胞系和肝癌细胞系,分别通过real-time PCR法和Western blotting法检测细胞中生物钟基因Clock、Bmal1、Per1、Per2、Per3、Cry1、Cry2、NPAS2的mRNA与蛋白。结果与转染AFP空载体L02细胞相比,AFP过表达的L02细胞中Clock的表达上调,Bmal1、Per1、Per2、Per3的表达下调(P均<0.05)。与转染AFP空载体SMMC-7721细胞相比,AFP过表达的SMMC-7721细胞中Clock的表达上调,Bmal1、Per1、Per2、Per3、Cry1的表达下调(P均<0.05)。结论 AFP可引起正常肝细胞和肝癌细胞生物钟基因表达紊乱,这可能是AFP促进肝癌细胞恶性生物学行为的原因之一。更多还原

【Abstract】 Objective To investigate the effect of alpha fetoprotein(AFP) on the expression of circadian clock genes in the normal hepatocytes and hepatoma cells.Methods The normal hepatocytes L02 and hepatocarcinoma cell line SMMC-7721 were infected with lentivirus vector with over-expressed AFP to construct the normal hepatocytes and hepatocarcinoma cell line with over-expression of AFP.The mRNA and protein expression levels of circadian clock genes such as Clock,Bmal1,Perl,Per2,Per3,Cry1,Cry2,and NPAS2 in the hepatocarcinoma cells before and after AFP infection were detected by real-time PCR and Western blotting,respectively.Results Compared with the L02 cells transfected with AFP empty vector,the expression of Clock was up-regulated,while the expression of Bmall,Per1,Per2,and Per3 was down-regulated in L02 cells with over-expression of AFP(all P<0.05).Compared with the SMMC-7721 cells transfected with AFP empty vector,the expression of Clock was up-regulated,while the expression of Bmall,Perl,Per2,Per3 and Cryl was down-regulated in the SMMC-7721 cells with over-expression of AFP(all P<0.05).Conclusion AFP can cause the disorder of circadian clock gene expression in the normal hepatocytes and hepatocarcinoma cells,which may be one of the reasons that AFP promotes the malignant biological behavior of hepatocarcinoma cells.更多还原