摘要
目的研究细胞周期检测点激酶1(Checkpoint kinase 1,Chk1)在胶质母细胞瘤细胞(Glioblastoma,GBM)中的表达及其与GBM细胞增殖、成瘤活性和预后生存的关联性。方法利用TCGA数据库和脑肿瘤分子数据库(Rembrandt)选择和分析GBM中Chk1的表达,并采用Western blot和Real-time PCR等分子生物学技术检测GBM细胞中Chk1的表达水平;通过慢病毒转染siRNA沉默Chk1的表达以探究其对于GBM细胞增殖和成球能力的影响;同时利用免疫组化染色及Rembrandt数据库对Chk1与GBM患者的预后生存关系进行分析。结果 TCGA和Rembrandt数据库分析结果表明Chk1在GBM组织中呈现高表达,同样Western blot和Real-time PCR检测结果显示Chk1在GBM细胞中呈现高表达;慢病毒转染siRNA特异性沉默Chk1能够显著抑制U87细胞的生长(P<0.01)和成球能力(P<0.05);预后生存分析结果显示Chk1低表达GBM患者的临床预后显著优于高表达患者(P<0.001)。结论 Chk1在GBM细胞中过量表达,表达上调能够促进GBM细胞的生长和增殖,并和GBM患者的不良预后相关。
Objective The aim of this study was to investigate the expression of cell cycle checkpoint kinase 1(Chk1)gene in glioblastoma cells(GBM)and its correlation with GBM cell proliferation,tumorigenic activity and prognosis.Methods The expression of Chk1 in GBM cells was selected and analyzed by TCGA database and brain tumor molecular database(Rembrandt),and the level of Chk1 expression in GBM cells was detected by molecular biology techniques such as Western blot and Real-Time PCR.The expression of Chk1 was silenced by siRNA to investigate its effect on proliferation and colony-forming ability of GBM cells.The prognosis survival of GBM patients accompanying with Chk1 expression was analyzed by immunohistochemical staining and Rembrandt database.Results The results of TCGA database and Rembrandt showed that Chk1 gene was highly expressed in GBM tissues.Western blot and Real-Time PCR also showed that Chk1 gene was highly expressed in GBM cells.Lentiviral transfection siRNA-specific silencing of Chk1 significantly inhibited proliferation and colony-forming ability of U87 cells(P<0.01 and P<0.05).Prognostic survival analysis showed that GBM patients with low expression of Chk1 gene had a significantly better clinical outcome than those of GBM patients with high expression of Chk1 gene(P<0.001).Conclusion Chk1 gene is overexpressed in GBM cells,up-regulation of Chk1 gene expression can promote the growth and proliferation of GBM cells,and Chk1 gene is associated with poor prognosis in GBM patients.
引文
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