TLR2对胶质母细胞瘤的预后价值和体外作用

Prognostic value and in vitro effect of TLR2 on glioblastoma multiforme

目的探究TCGA数据库中TLR2基因在胶质母细胞瘤(GBM)中的预后价值,并明确该基因对胶质母细胞瘤细胞LN229的体外作用。方法从TCGA数据库中获取169例胶质母细胞瘤患者的临床信息和肿瘤样本的转录组数据,通过Kaplan-Meier生存曲线分析评估TLR2基因在GBM患者中的预后作用。构建TLR2敲减的人胶质母细胞瘤LN229细胞系,通过RT-qPCR和Western Blot验证。通过CCK8、克隆形成、细胞划痕、Transwell小室法,检测敲减TLR2对LN229细胞的增殖、克隆形成、迁移、侵袭能力的影响。结果与正常样本相比,TLR2在GBM患者样本中表达增高。生存分析结果显示,TLR2高表达组患者总生存期显著低于低表达组患者(总生存期中位值333 d vs.402 d,P<0.05)。RT-qPCR和Western Blot结果显示,TLR2敲减的人胶质母细胞瘤LN229细胞系成功构建。与转染si-control的LN229胶质瘤细胞相比,转染si-TLR2的LN229胶质瘤细胞增殖能力明显降低(P<0.05)。与转染si-control的LN229胶质瘤细胞相比,转染si-TLR2的LN229胶质瘤细胞克隆形成数明显降低(P<0.05)。转染si-TLR2的LN229细胞迁移能力明显低于转染si-control的LN229胶质瘤细胞(P<0.05)。转染si-TLR2的LN229细胞侵袭能力明显低于转染si-control的LN229胶质瘤细胞(P<0.05)。结论本研究通过数据挖掘发现TLR2表达与胶质母细胞瘤患者的预后显著相关,通过实验验证敲减TLR2有效抑制LN229细胞的增殖、迁移、侵袭和克隆形成能力,上述发现将为胶质母细胞瘤预后判断和靶向治疗提供新的潜在靶点。

Objective To explore the prognostic value of TLR2 gene in TCGA database in glioblastoma Multiforme(GBM),and clarify the in vitro effect of TLR2 gene on glioblastoma multiforme cell LN229.Methods The clinical information of 169 patients with glioblastoma multiforme and the transcriptome data of tumor samples were obtained from the TCGA database.The prognostic effect of TLR2 gene in GBM patients was evaluated by Kaplan-Meier survival curve analysis.TLR2 was knocked down by small interfering RNA in LN229,and the knockdown efficiency of TLR2 was verified by RT-qPCR and Western Blot.The effects of TLR2 on the proliferation,clone formation,migration,and invasion of LN229 cells were detected by CCK8,clone formation,cell scratch and Transwell chamber.Results Compared with normal samples,TLR2 expression was increased in GBM patients.The overall survival of patients in the high expression group was lower than that in the low expression group(overall survival 333 days vs.402 days,P<0.05).The results of RT-qPCR and Western Blot showed that TLR2 was well inhibited in TLR2-kockdowned LN229.The results of CCK8 assay showed that Human glioblastoma LN229 cell line with TLR2 knockdown was successfully constructed.Compared with LN229 glioma cells transfected with si-control,the number of clone formation of LN229 glioma cells transfected with si-TLR2 was significantly reduced(P<0.05).The migration ability of LN229 cells transfected with si-TLR2 was significantly lower than that of LN229 glioma cells transfected with si-control(P<0.05).The invasion ability of LN229 transfected with si-TLR2 was significantly lower than that of LN229 transfected with si-control(P<0.05).Conclusions In this study,data analysis showed that TLR2 expression was significantly correlated with the prognosis of patients with glioblastoma multiforme.Knockdown of TLR2 in vitro effectively inhibited the proliferation,migration,invasion and colony formation of LN229 cells.Our finding will provide a new potential target for prognosis judgment and targeted therapy of glioblastoma multiforme.