着丝粒蛋白U促进肝癌细胞增殖能力的实验研究

Experimental study on Centromere Protein-U in promoting proliferation of hepatocellular carcinoma cells

目的研究CENPU在肝癌中的表达情况,进一步研究CENPU对肝癌细胞增殖能力的影响。方法通过GEPIA数据库和GEO数据库分析CENPU在肝癌组织和癌旁组织中CENPU的表达情况。对TCGA数据库中370例肝癌患者的临床信息进行分析并绘制生存曲线,评估CENPU的预后价值。利用UALCAN在线数据库分析CENPU对肝癌患者预后的影响。RT-qPCR方法检测3种肝癌细胞和1种人正常肝细胞系LO2中CENPU的表达情况。免疫组织化学染色检测本中心10例肝癌组织与癌旁组织标本中CENPU的表达情况。CCK8法检测CENPU对肝癌细胞增殖能力的影响。裸鼠移植瘤实验评估CENPU对肝癌细胞体内成瘤能力的影响。结果GEPIA数据库分析结果显示肝癌组织中CENPU表达量明显高于癌旁组织(P<0.05)。GSE29721、GSE45514和GSE84405数据集中肝癌组织中CENPU表达量均显著高于癌旁组织(P<0.01,P<0.001,P<0.001)。TCGA数据库肝癌数据集的生存分析结果显示,高表达CENPU的肝癌患者总体生存期短于低表达CENPU的肝癌患者。UACLAN在线数据库分析结果提示高表达CENPU的肝癌患者预后更差(P<0.05)。RT-qPCR检测结果显示3种人肝癌细胞系中CENPU mRNA表达量均高于人正常肝细胞系。10例癌旁组织中的CENPU表达量均低于肝癌组织。CCK8实验结果显示敲低CENPU后肝癌细胞的增殖能力明显受到抑制。裸鼠移植瘤实验结果提示,敲低CENPU后肝癌细胞的成瘤能力明显降低。结论CENPU在肝癌中高表达且与不良预后相关,CENPU促进肝癌细胞的增殖能力。

Objective To study the expression of Centromere Protein-U(CENPU)in hepatocellular carcinoma,and further investigate the effect of CENPU on the proliferative ability of hepatocellular carcinoma.Methods The expression of CENPU in hepatocellular carcinoma tissues and para-cancerous tissues was analyzed by GEPIA database and GEO database.Clinical information of 370 patients suffering from hepatocellular carcinoma from the TCGA database was analyzed and survival curves were plotted to assess the prognostic value of CENPU.The effect of CENPU on the prognosis of patients with hepatocellular carcinoma was analyzed by using the UALCAN online database.RT-qPCR method was used to detect the expression of CENPU in three types of hepatocellular carcinoma cells and one human normal liver cell line LO2.Immunohistochemical staining was used to detect the expression of CENPU in 10 specimens of hepatocellular carcinoma tissues and para-cancerous tissues from our center.CCK8 method was used to detect the effect of CENPU on the proliferative ability of hepatocellular carcinoma cells.Nude mice transplantation tumor assay was performed to evaluate the effect of CENPU on the tumor-forming ability of hepatocellular carcinoma cells in vivo.Results The analysis of GEPIA database showed that the expression of CENPU in hepatocellular carcinoma tissues was significantly higher than that in para-cancerous tissues(P<0.05).The expression of CENPU in hepatocellular carcinoma tissues was higher than that in para-cancerous tissues in the datasets of GSE29721,GSE45514,and GSE84405(P<0.01,P<0.001,P<0.001).The survival analysis results of the hepatocellular carcinoma data in TCGA database showed that the overall survival of hepatocellular carcinoma patients with high CENPU expression was shorter than that of hepatocellular carcinoma patients with low CENPU expression.The analysis results of the UACLAN online database suggested that the prognosis of hepatocellular carcinoma patients with high CENPU expression was worse(P<0.05).RT-qPCR results showed that the mRNA expression of CENPU in the 3 human hepatocellular carcinoma cell lines were higher than that in normal liver cell lines.The expression of CENPU in 10 cases of para-neoplastic tissues was lower than that in hepatocellular carcinoma tissues.The proliferation of hepatocellular carcinoma cells was significantly inhibited after knockdown of CENPU in CCK8 assay.The results of nude mice transplantation tumor assay suggested that the tumorigenic ability of hepatocellular carcinoma cells was significantly reduced after knocking down CENPU.Conclusions CENPU is high expressed in hepatocellular carcinoma,which is associated with poor prognosis,and CENPU promotes the proliferation ability of hepatocellular carcinoma cells.