CRY2过表达对肝癌细胞增殖及生物钟基因、蛋白表达的影响  

Effects of CRY2 overexpression on cell proliferation, clock gene and protein expression of hepatocellular carcinoma cells

作　　者：许静 孙诚谊[1,2] 喻超 何晓晓[3] 杨盛力 XU Jing;SUEN Chengyi;YU Chao;HE Xiaoxiao;YANG Shengli(Guizhou Medical University, Guiyang 550000, China)

机构地区：[1]贵州医科大学,贵阳550000 [2]贵州医科大学肝胆胰脾重点实验室 [3]华中科技大学同济医学院附属梨园医院

出　　处：《山东医药》2019年第8期5-8,共4页Shandong Medical Journal

基　　金：贵州省科学技术基金项目(黔科合J字2015-2013);贵州省科学技术厅-贵州医科大学附属医院联合基金(黔科合LH字2016-7229)

摘　　要：目的观察CRY2过表达对肝癌细胞系HepG2增殖及生物钟基因、蛋白表达的影响。方法将HepG2细胞分为实验组和对照组,分别转染CRY2过表达病毒上清液及阴性对照病毒上清液,以未转染的HepG2细胞为空白组,分别于转染24、48、72、96 h后采用CCK-8实验观察细胞增殖能力变化。采用real-time PCR法检测实验组和对照组细胞中的生物钟基因CLOCK、BMAL1、PER1、PER2、PER3、DEC1、DEC2、CRY1、CRY2、NPAS2,采用Western blotting法检测上述生物钟基因蛋白。结果转染48、72、96 h后,实验组细胞增殖能力较对照组和空白组降低(P均<0.05)。实验组细胞中PER1、PER2、PER3、CLOCK、CRY1、CRY2、BMAL1、NPAS2、DEC2 mRNA及蛋白相对表达量均高于对照组,DEC1 mRNA及蛋白相对表达量均低于对照组(P均<0.05)。结论 CRY2过表达可抑制肝癌细胞增殖,同时在mRNA及蛋白水平上调相关生物钟基因表达。CRY2表达上调后可能通过调控生物钟网络从而抑制肝癌细胞的增殖能力。Objective To investigate the effects of overexpression of circadian clock gene CRY2 on the cell proliferation, clock gene and protein expression of hepatocellular carcinoma cells HepG2. Methods HepG2 cells were divided into the experimental group and control group, CRY2 overexpressing virus supernatant and negative control virus supernatant were transfected, respectively, and meanwhile, the untransfected HepG2 cells were used as the blank group. The CCK8 assay was used to detect the proliferation of hepatoma cells at 24, 48, 72, and 96 h after transfection, and the real-time PCR was used to detect the mRNA expression of the clock genes CLOCK, BMAL1, PER1, PER2, PER3, DEC1, DEC2, CRY1, CRY2, NPAS2, and Western blotting was used to detect the protein expression of the circadian clock gene. Results At 48, 72, and 96 h after transfection, the cell proliferation ability of the experimental group was lower than those of the control group and the blank group (all P <0.05). The relative mRNA and protein expression levels of PER1, PER2, PER3, CLOCK, CRY1, CRY2, BMAL1, NPAS2, and DEC2 were higher in the experimental group than in the control group, and the relative mRNA and protein expression levels of DEC1 were lower than those in the control group (all P < 0.05 ).Conclusion The overexpression of CRY2 can inhibit the proliferation of hepatoma cells, and up-regulate the expression of relevant clock genes in the mRNA and protein levels, indicating that CRY2 plays an important regulatory role in the circadian clock gene network.

关 键 词：肝细胞癌 生物钟 生物钟基因 CRY2 细胞增殖 

分 类 号：R735.7[医药卫生—肿瘤]