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基于TCGA数据库和实验验证分析肝细胞癌中PLCβ4基因的表达及临床意义

Expression and Clinical Significance of PLCβ4 Gene in Hepatocellular Carcinoma Analyzed Based on TCGA Database and Experimental Validation

  • 摘要:
    目的 基于癌症基因组图谱(TCGA)数据库分析肝细胞癌中PL-Cβ4基因mRNA的表达及其临床意义。
    方法 基于TCGA数据库中的424例临床样本资料(包括肝细胞癌组织374例,非肿瘤肝组织50例),采用Kaplan-Meier法、Cox回归分析、免疫浸润分析,评估PLCβ4基因与HCC患者临床特征及生存预后的关系。应用PLCβ4基因与24种免疫细胞的相关性分析,观察PLCβ4基因与免疫细胞浸润的关系及与肝癌高频突变基因TP53 mRNA表达水平的相关性。另外收集新疆医科大学第一附属医院病理科HCC患者的高、中、低分化肿瘤组织以及正常肝组织石蜡切片各20例,HE染色法进行组织病理学观察,免疫组织化学法对各临床样本中PLCβ4、Ki-67蛋白表达水平进行验证。
    结果  PLCβ4基因在HCC中的表达水平显著高于正常组织(P<0.01),且PLCβ4高表达患者的总生存期均明显长于低表达者(P<0.05),提示PLCβ4显著影响HCC患者预后。相关性分析结果显示PLCβ4基因的表达水平与免疫细胞浸润高度相关;且PLCβ4与TP53基因的表达水平高度相关。HCC组织样本中的PLCβ4基因表达显著高于正常组织(P<0.001),且与分化程度呈负相关关系。
    结论 PLCβ4可作为肝细胞癌的一个独立预后因素,有望成为HCC治疗的新型分子靶点。

     

    Abstract:
    Objective To analyze the PLCβ4 gene mRNA expression and its clinical significance in hepatocellular carcinoma (HCC) based on TCGA database.
    Methods Based on the data on 424 clinical samples (including 374 cases of HCC tissues and 50 cases of nontumor liver tissues) in the TCGA database, Kaplan–Meier method, Cox regression analysis, and immune infiltration analysis were performed to evaluate the relationship between PLCβ4 gene and the clinical characteristics and survival prognosis of HCC patients. Correlation analysis between PLCβ4 gene and 24 types of immune cells was applied to investigate the relationship between PLCβ4 gene and immune cell infiltration and mRNA expression level of TP53 gene, a high-frequency mutation gene in HCC. In addition, paraffin sections of highly, moderately, and poorly differentiated tumor tissues and normal liver tissues from HCC patients were collected. The histopathological observation was carried out via HE staining method, and the expression levels of PLCβ4 and Ki-67 proteins in each clinical sample were verified through the immunohistochemical method.
    Results The expression level of PLCβ4 gene in HCC was significantly higher than that in normal tissues (P<0.01), and all patients in the PLCβ4 high-expression group had a significantly longer overall survival than those in the low-expression group (P<0.05), which suggested that PLCβ4 substantially affected the prognosis of HCC patients. Correlation analysis showed that the expression level of PLCβ4 gene was highly correlated with immune cell infiltration and the expression level of TP53 gene. As verified by clinical sample experiments, HE staining experiments and immunohistochemical results revealed that PLCβ4 gene expression in HCC tissue samples was significantly higher than that in normal tissues (P<0.001), and it was negatively correlated with the degree of differentiation.
    Conclusion PLCβ4 may serve as an independent prognostic factor in HCC and is expected to be a novel molecular target for HCC treatment.

     

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