Li Ling, Huang Kang, Zhao Huijia, Chen Binyao, Ye Qifa, Yue Jiang
Zhongnan Hospital of Wuhan University, Institute of Hepatobiliary Diseases of Wuhan University, Transplant Center of Wuhan University, Hubei Key Laboratory of Medical Technology on Transplantation, Wuhan, Hubei, RP China.
Department of Pharmacology, Basic Medical School of Wuhan University, Wuhan, Hubei, RP China.
Cell Cycle. 2020 May;19(10):1236-1252. doi: 10.1080/15384101.2020.1749471. Epub 2020 Apr 10.
This study aims to investigate the potential mechanisms and identify core biomarkers of Hepatocellular carcinoma (HCC). The profile GSE113850 was downloaded to analyze the differentially expressed genes. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and protein-protein interaction network analysis were used to reveal the main signal pathways of the differentially expressed genes (DEGs) and hub genes. The correlation between core gene expression and pathological stages, the disease-free survival analysis, the overall survival analysis were analyzed by Gene Expression Profiling Interactive Analysis. Furthermore, we reidentified the expression level of core genes of carcinoma tissues and para-carcinoma tissues from 14 HCC patients with real-time reverse transcription-polymerase chain reaction analysis (RT-PCR) and western blotting. After SK-Hep1 cell was treated with cyclin-dependent kinase 1 (CDK1) siRNA for 72 h, we detected the expression of the core genes and fluorescence-activated cell sorting analysis. A total of 378 DEGs were found. GO and KEGG analysis revealed that the DEGs were mainly enriched in the cell cycle. There were positive correlations among CDK1, polo-like kinase 1, shugoshin2 and anillin actin-binding protein. Moreover, the expression levels of four core genes were related to the HCC occurrence, pathological stages, and survivorship curve. The clinical HCC specimens verified the higher expression level of core genes by real-time RT-PCR. The transfection of siCDK1 in SK-Hep1 resulted in a disordered cell cycle. Furthermore, CDK1 knockdown suppressed the expression of PLK1, ANLN, and SGOL2. The CDK1-PLK1/SGOL2/ANLN pathway mediating abnormal cell division in the cell cycle might be a critical process in HCC.
本研究旨在探究肝细胞癌(HCC)的潜在机制并鉴定其核心生物标志物。下载基因表达谱GSE113850以分析差异表达基因。采用基因本体论(GO)、京都基因与基因组百科全书(KEGG)以及蛋白质-蛋白质相互作用网络分析来揭示差异表达基因(DEGs)和枢纽基因的主要信号通路。通过基因表达谱交互式分析来分析核心基因表达与病理分期之间的相关性、无病生存分析以及总生存分析。此外,我们通过实时逆转录-聚合酶链反应分析(RT-PCR)和蛋白质免疫印迹法重新鉴定了14例HCC患者癌组织和癌旁组织中核心基因的表达水平。用细胞周期蛋白依赖性激酶1(CDK1)小干扰RNA(siRNA)处理SK-Hep1细胞72小时后,我们检测了核心基因的表达并进行了荧光激活细胞分选分析。共发现378个差异表达基因。GO和KEGG分析显示,差异表达基因主要富集于细胞周期。CDK1、polo样激酶1、shugoshin2和肌动蛋白结合蛋白anillin之间存在正相关。此外,四个核心基因的表达水平与HCC的发生、病理分期及生存曲线相关。临床HCC标本通过实时RT-PCR验证了核心基因的高表达水平。在SK-Hep1细胞中转染siCDK1导致细胞周期紊乱。此外,敲低CDK1可抑制PLK1、ANLN和SGOL2的表达。细胞周期中由CDK1-PLK1/SGOL2/ANLN途径介导的异常细胞分裂可能是HCC中的一个关键过程。