基质金属蛋白酶7在肝癌细胞迁移及免疫细胞浸润中的作用与预后价值  

Role and prognostic value of matrix metalloproteinase-7 in the migration and immune cell infiltration of hepatocellular carcinoma

作　　者：刘淑岩 杨其昌[1] 沈屹[1] 周虹 钱金锋[1] LIU Shuyan;YANG Qichang;SHEN Yi;ZHOU Hong;QIAN Jinfeng(Department of Pathology,The First People’s Hospital of Nantong,Nantong,Jiangsu 226014,China)

机构地区：[1]南通市第一人民医院病理科,江苏南通226014

出　　处：《临床肝胆病杂志》2024年第7期1420-1427,共8页Journal of Clinical Hepatology

基　　金：南通大学研究生创新计划项目(YKC16070)。

摘　　要：目的评估基质金属蛋白酶7(MMP7)在肝癌细胞迁移及免疫细胞浸润中的作用及预后价值。方法分别构建下调或上调靶基因MMP7的MMP7_siRNA、pMMP7转染肝癌细胞株(MHCC97H)。采用RT-qPCR、Western Blot分别检测细胞中靶基因mRNA及蛋白的表达水平。扫描电镜和Transwell小室实验分别观察细胞伪足和迁移能力的变化,并采用生物信息学的方法,在TCGA、TIMER数据库分析MMP7与免疫细胞及肝癌患者免疫浸润评分之间的相关性,并进一步研究MMP7与肝癌患者预后的相关性。相关性分析采用Spearman方法。Sanger Box在线工具评估MMP7在肝癌总体生存期、疾病特异性生存期方面的意义。Kaplan-Meier法绘制生存曲线,Log-rank检验评估不同组样本之间的预后差异。结果构建的MMP7_siRNA、pMMP7转染MHCC97H细胞后均能有效下调或上调靶基因MMP7的表达,在MHCC97H细胞中MMP7被干扰后细胞伪足明显减少,并且变短,但是过表达MMP7之后细胞表面丝状伪足数量明显增多,并且伪足长度变长,放射状排列。Transwell小室结果发现,MMP7_siRNA2能够显著降低细胞迁移能力(P<0.05),而转染pMMP7后,细胞迁移能力显著增加(P<0.05)。MMP7的表达与B淋巴细胞(r=0.37)、CD4+T淋巴细胞(r=0.40)、中性粒细胞(r=0.49)、巨噬细胞(r=0.49)、树突状细胞(r=0.47)显著相关(P值均<0.05)。在TCGA数据库中,基于总体生存期最佳截断值将肝癌患者分成MMP7高表达组(n=267)和MMP7低表达组(n=146),结果发现,MMP7高表达组的总体生存期明显低于MMP7低表达组(P<0.05);基于疾病特异性生存期最佳截断值将肝癌患者分成MMP7高表达组(n=257)和MMP7低表达组(n=145),结果发现,MMP7高表达组的疾病特异性生存期也低于MMP7低表达组(P<0.05)。结论MMP7促进了肝癌细胞的迁移,并在免疫细胞浸润中发挥主要作用,MMP7的表达也与肝癌的预后具有明显相关性。Objective To investigate the role and prognostic value of matrix metalloproteinase-7(MMP7)in the migration and immune cell infiltration of hepatocellular carcinoma.Methods MMP7_siRNA for downregulating the target gene MMP7 and pMMP7 for upregulating MMP7 were constructed and were used to transfect hepatocellular carcinoma cell line(MHCC97H).RT-qPCR and Western Blot were used to measure the mRNA and protein expression levels of the target gene in cells.Scanning electron microscopy and Transwell assay were used to observe the changes in cell pseudopodia and migration ability,and bioinformatics methods were used to investigate the correlation of MMP7 with immune cells and immune infiltration score in TCGA and TIMER databases in patients with hepatocellular carcinoma,as well as the association between MMP7 and the prognosis of patients with hepatocellular carcinoma.The Spearman method was used for correlation analysis.Sanger Box online tool was used to assess the value of MMP7 in the overall survival curve and disease-specific survival of hepatocellular carcinoma.The Kaplan-Meier method was used to plot survival curves,and the Log-rank test was used for comparison of prognosis between different samples.Results After MHCC97H cells were transfected with MMP7_siRNA or pMMP7,there was a significant reduction or increase in the expression of the target gene MMP7;after downregulation of MMP7,there were significant reductions in the number and length of the pseudopodia,while after MMP7 overexpression,there were significant increases in the number and length of filopodia with radial arrangement.The Transwell chamber assay showed that MMP7_siRNA2 significantly reduced the migration ability of cells(P<0.05),and there was a significant increase in migration ability after pMMP7 transfection.The expression of MMP7 was significantly correlated with B lymphocytes(r=0.37,P<0.05),CD4+T lymphocytes(r=0.40,P<0.05),neutrophils(r=0.49,P<0.05),macrophages(r=0.49,P<0.05),and dendritic cells(r=0.47,P<0.05).In the TCGA database,the patients

关 键 词：肝肿瘤 基质金属蛋白酶7 细胞运动 

分 类 号：R735.7[医药卫生—肿瘤]