胶质瘤GPC6表达水平与患者预后的关系以及体外GPC6对胶质瘤细胞增殖的影响  

Correlation of GPC6 expression level in gliomas with prognosis of patients and effect of GPC6 on proliferation of glioma cells in vitro

作　　者：延冰 张坤 徐心怡 刘华 Yan Bing;Zhang Kun;Xu Xinyi;Liu Hua(Department of General Surgery,Central Hospital Affiliated to Shandong First Medical University,Jinan 250013,China;Cell Therapy Center,Central Hospital Affiliated to Shandong First Medical University,Jinan 250013,China;Institute of Environmental Science,College of Resources and Environment,Shandong Agriculture University,Taian 271018,China)

机构地区：[1]山东第一医科大学附属中心医院普通外科,济南250013 [2]山东第一医科大学附属中心医院细胞治疗中心,济南250013 [3]山东农业大学资源与环境学院环境生态工程系,泰安271018

出　　处：《肿瘤研究与临床》2025年第3期198-205,共8页Cancer Research and Clinic

摘　　要：目的探讨磷脂酰肌醇蛋白聚糖6(GPC6)在胶质瘤中的表达水平与患者预后的相关性,以及体外GPC6对胶质瘤细胞增殖的影响。方法从癌症基因组图谱(TCGA)数据库中下载胶质瘤患者GPC6基因转录组测序(RNA⁃seq)数据及临床资料,入组667例,以入组患者GPC6 RNA⁃seq的中位FPKM值区分GPC6基因的低表达和高表达,采用Kaplan⁃Meier法分析有生存数据的664患者中GPC6高、低表达组总生存(OS),组间比较采用log⁃rank检验。取回顾性收集的2008年2月至2011年10月手术胶质瘤患者术后肿瘤组织芯片,采用免疫组织化学(IHC)法检测147例样本中GPC6蛋白的表达情况,按阳性细胞比例与染色强度相结合的半定量评分是否≥4分区分GPC6蛋白高表达和低表达。比较147例患者临床病理特征分层组间肿瘤组织GPC6蛋白高、低表达患者的分布,采用Kaplan⁃Meier法分析GPC6蛋白高、低表达组患者OS和无病生存(DFS),采用单因素和多因素Cox比例风险模型分析影响患者OS和DFS不良的危险因素。用包装有GPC6小干扰RNA的慢病毒、包装有无关对照序列的慢病毒感染人胶质瘤U251细胞,分别为ShGPC6组和ShCtrl组;每天计数绿色荧光细胞并计算细胞增殖倍数,共5 d,绘制细胞增殖曲线。结果TCGA数据库中,667例胶质瘤患者中位FPKM值为225.66,世界卫生组织(WHO)分类低级别胶质瘤患者(515例)GPC6基因中位FPKM值低于高级别胶质瘤患者(152例)(201.10比347.92),差异有统计学意义(Z=5.36,P<0.001);GPC6低表达组(332例)患者OS优于高表达组(332例),差异有统计学意义(P<0.001)。对组织芯片IHC染色显示,GPC6蛋白在胶质瘤细胞中主要表达于细胞质和细胞核中;GPC6蛋白高表达患者占31.3%(46/147),死亡、复发、高级别、生存时间≤84个月、表皮生长因子受体阳性患者中GPC6蛋白高表达患者占比均较高,差异均有统计学意义(均P<0.05),不同性别、年龄是否≤43岁、Ki⁃67是否阳性和程�Objective To investigate the correlation between the expression level of glypican-6(GPC6)in gliomas and the prognosis of patients,as well as the effect of GPC6 on the proliferation of glioma cells in vitro.Methods The transcriptome sequencing(RNA-seq)data of GPC6 gene and clinical data of glioma patients were downloaded from The Cancer Genome Atlas(TCGA)database.A total of 667 patients were enrolled,and the median FPKM value of GPC6 RNA-seq was used to distinguish the low and high expression of GPC6 gene.Kaplan-Meier method was used to analyze the overall survival(OS)of GPC6 high and low expression groups in 664 patients with survival data,and log-rank test was used for inter group comparison.The postoperative tumor tissue chips were retrospectively collected from glioma patients who underwent surgery from February 2008 to October 2011,immunohistochemistry(IHC)was used to detect the expression of GPC6 protein in 147 samples,and semi-quantitative scoring≥4 points or not was used to distinguish the high and low expression of GPC6 protein based on the proportion of positive cells and staining intensity.The distributions of patients with high and low expression of GPC6 protein in tumor tissues were compared between different clinical pathological features in 147 patients,and Kaplan-Meier method was used to analyze the overall survival and disease-free survival(DFS)of patients in the GPC6 protein high and low expression groups;the risk factors that affect poor OS and DFS in patients were analyzed using univariate and multivariate Cox proportional hazards models.The human glioma U251 cells were infected with lentivirus packaged with GPC6 small interfering RNA and lentivirus packaged with unrelated control sequences,namely ShGPC6 group and ShCtrl group,respectively;green fluorescent cells were counted daily and the cell proliferation fold was calculated for 5 days,and the cell proliferation curve was plotted.Results In TCGA database,the median FPKM value of 667 glioma patients was 225.66.The median FPKM value of GPC6

关 键 词：胶质瘤 磷脂酰肌醇蛋白聚糖类 组织阵列分析 预后 免疫组织化学 细胞增殖 

分 类 号：R739.4[医药卫生—肿瘤]