CCL18在胶质母细胞瘤中的表达及其对U87MG细胞增殖和迁移的影响

Expression of CCL18 in glioblastoma and its effect on proliferation and migration of human U87MG cells

目的探讨趋化因子配体18(CCL18)在胶质母细胞瘤(GBM)中的表达与GBM临床预后的关系以及对U87MG细胞增殖和迁移的影响。方法基于癌症基因组图谱(TCGA)和基因型-组织表达(GTEx)数据库中GBM和正常脑组织数据,分析CCL18在GBM与正常脑组织中的表达差异,采用免疫组织化学染色进行验证。应用Kaplan-Meier生存分析、单因素和多因素Cox回归分析评估CCL18对GBM患者预后的影响。基于多因素Cox回归模型构建生存列线图,绘制校准曲线和受试者工作特征(ROC)曲线进行验证。将人胶质母细胞瘤U87MG细胞分为siRNACCL18组、siRNA-NC组和Mock-siRNA组,采用实时荧光定量PCR(qPCR)法检测细胞中CCL18 mRNA表达,CCK-8法和细胞划痕愈合实验检测U87MG细胞的增殖和迁移能力。结果TCGA和GTEx数据库分析结果显示,GBM组织中CCL18基因表达水平高于正常脑组织;免疫组织化学染色结果显示,GBM组CCL18阳性表达率高于对照组(P<0.05)。CCL18高表达组患者预后较低表达组差(P<0.01)。Cox回归分析结果显示,IDH野生型和CCL18高表达是GBM患者死亡的独立危险因素(P<0.05)。校准曲线显示列线图预测模型与实际生存有较好一致性,ROC曲线表明该模型对GBM患者生存概率有较好预测能力。siRNA-CCL18组CCL18 mRNA表达水平、细胞增殖(48 h、72 h和96 h)和迁移能力(24 h)均低于siRNA-NC组和Mock-siRNA组(P<0.05)。结论CCL18在GBM中表达上调,可促进GBM肿瘤细胞增殖和迁移,且可导致患者预后不良。

Objective To analyze the expression chemokine(C-C motif)ligand 18(CCL18)on the clinical prognosis of glioblastoma(GBM)and its effects on the proliferation and migration of U87MG cells.Methods The differential expression of CCL18 gene in GBM and normal brain tissue was analyzed based on the GBM data from Cancer Genome Atlas(TGGA)database and the normal brain tissue data from Genotype-Tissue Expression(GTEx)database.Immunohistochemical staining was used to verification.Kaplan-Meier survival analysis,univariate and multivariable Cox regression analysis were used to evaluate the impact of CCL18 expression on the prognosis in GBM patients.A nomogram was established based on the results of multivariate Cox analysis.The calibration curve and receiver-operating characteristic(ROC)curve were draw for validation.Human glioblastoma U87MG cells were divided into the siRNA-CCL18 group,the siRNA-NC group and the Mock-siRNA group.qPCR was used to detect the content of CCL18 mRNA in the different U87MG cell groups.The CCK-8 assay and cell scratch test were used to detect the cell proliferation and migration ability of U87MG cells in each group.Results CCL18 was significantly upregulated in GBM tissue(P<0.05).The immunohistochemistry results suggested that CCL18 protein was significantly increased in GBM tissue(P<0.05).Survival analysis indicated that high expression of CCL18 was associated with poor prognosis in GBM patients(P<0.05).Multivariate Cox regression analysis revealed that IDH wild-type and high level of CCL18 expression were the two major risk factors affecting the poor prognosis of GBM(P<0.05).The calibration curve showed that the actual survival rate of patients was consistent with the predicted survival rate.The ROC curve demonstrated that the model had a good predictive for predicting the survival of GBM patients.siRNA reduced the expression of CCL18 in human U87MG cells.The results of CCK-8 assay showed that the proliferation of U87MG cells was significantly inhibited(P<0.05),and the wound healing scratch ability of U87MG cells with low expression of CCL18 decreased(P<0.01).Conclusion CCL18 is over-expressed in GBM tissue,and high CCL18 expression is associated with poor prognosis in GBM patients.CCL18 promotes GBM cell proliferation and migration and may act as a relevant predictive biomarker for GBM.