摘要
目的比较肝癌复发与预后良好患者癌组织的蛋白表达,分析miR-152-3p相关的靶蛋白差异及调控机制,探讨miR-152-3p在肝癌复发中的作用。方法应用TMT标记全蛋白质组学测序方法和RT-PCR分别检测肝癌切除术后2年复发(n=6)与5年预后良好(n=6)患者肝癌组织的蛋白质表达和miR-152-3p的表达水平;联合六大数据库检索分析miR-152-3p靶基因,并运用Gene Ontology、DAVID和REACTOME数据库进行靶基因筛选、富集注释和信号转导通路富集分析;将筛选的miR-152-3p靶基因进行基因突变频率和生存曲线分析,验证miR-152-3p靶基因在肝癌复发中的作用。计量资料2组间比较采用独立样本t检验。肝脏组织有关基因生存率分析采用Kaplan-Meier分析。结果肝癌切除术后预后良好患者癌组织的miR-152-3p转录表达水平显著高于复发患者癌组织的水平(P<0.05)。蛋白测序结果显示,预后良好患者与复发患者的癌组织存在365个差异表达蛋白,联合肝癌复发数据库分析显示,其中有17个蛋白受miR-152-3p调控。进一步的信号通路分析发现,17个受miR-152-3p调控的靶基因其功能集中于线粒体核糖体翻译调控;多重富集发现靶基因与线粒体呼吸链复合体密切关系的蛋白6个,分别为AKAP1、FOXRED1、MRPL28、MRPL50、SHC1、STAU1。基因突变频率及生存曲线分析发现,线粒体呼吸链相关靶蛋白的功能缺失或减弱会严重影响患者的生存率。结论miR-152-3p在HCC切除术后预后良好和复发患者的癌组织表达差异显著,miR-152-3p在肝癌复发中作用机制可能是通过调控靶基因AKAP1、FOXRED1、MRPL28、MRPL50、SHC1、STAU1作用于线粒体呼吸链,影响细胞氧化呼吸功能导致肝癌复发。
Objective To investigate the difference in protein expression between hepatocellular carcinoma(HCC)patients with recurrence and those with good prognosis,the differential expression and regulatory mechanism of miR-152-3p target proteins,and the role of miR-152-3p in the recurrence of HCC.Methods TMT-labeled proteomic sequencing and RT-PCR were used to measure the expression of proteins and the expression of miR-152-3p in the HCC tissue of six patients with recurrence at 2 years after HCC resection and six patients with good prognosis at 5 years.Six databases were used to analyze the target genes of miR-152-3p,and Gene Ontology,DAVID,and REACTOME databases were used to perform target gene screening,enrichment annotation,and signal transduction pathway enrichment analysis.Gene mutation frequency and survival curve analysis were performed for the target genes of miR-152-3p to verify the role of miR-152-3p target genes in patients with HCC recurrence.The independent samples t-test was used for comparison of continuous data between two groups,and a Kaplan-Meier analysis was performed to investigate the survival rates of liver-related genes.Results Compared with the patients with HCC recurrence,the patients with good prognosis after HCC resection had a significantly higher transcriptional expression level of miR-152-3p in HCC tissue(P<0.05).The results of protein sequencing showed that there were 365 differentially expressed proteins in HCC tissue between the patients with good prognosis and the patients with recurrence,and the analysis of HCC recurrence databases showed that 17 proteins were regulated by miR-152-3p.Further analysis of the signaling pathways showed that the function of the 17 target genes regulated by miR-152-3p was enriched in the translation and regulation of mitochondria and ribosome,and multiple enrichment revealed that six target genes were closely associated with mitochondrial respiratory chain complex,i.e.,AKAP1,FOXRED1,MRPL28,MRPL50,SHC1,and STAU1.Gene mutation frequency and survival curve analysis showed that the loss or weakening of the function of mitochondrial respiratory chain-related target proteins seriously affected the prognosis and survival rate of patients.Conclusion There is a significant difference in the expression of miR-152-3p in HCC tissue between patients with good prognosis and those with recurrence after HCC resection,and miR-152-3p may lead to the recurrence of HCC by regulating the target genes AKAP1,FOXRED1,MRPL28,MRPL50,SHC1,and STAU1,acting on the mitochondrial respiratory chain,and affecting the oxidative respiratory function of cells.
作者
刘晨霞
常凯
那琬琳
王艳艳
牟东
李华
江忠勇
刘媛
熊杰
LIU Chenxia;CHANG Kai;NA Wanlin;WANG Yanyan;MOU Dong;LI Hua;JIANG Zhongyong;LIU Yuan;XIONG Jie(School of Clinical Medicine,Southwest Medical University,Luzhou,Sichuan 646000,China;Department of Clinical Laboratory,The General Hospital of Western Theater Command,Chengdu 610083,China;Department of Gastroenterology,The General Hospital of Western Theater Command,Chengdu 610083,China;Department of Oncology,The General Hospital of Western Theater Command,Chengdu 610083,China)
出处
《临床肝胆病杂志》
CAS
北大核心
2021年第2期364-369,共6页
Journal of Clinical Hepatology
基金
国家自然科学基金(81301445)
卫生部应用研究项目(28-1-16)。
