目的:评估不同阶段结肠腺癌患者中CDC样激酶2 (CLK2)基因的表达,以及CLK2在结肠腺癌细胞中的表达变化和CLK2所介导的细胞功能。方法:在TIMER数据库和GEPIA数据库分析CLK2在健康与不同肿瘤组织中的表达水平,找到差异性表达显著的肿瘤类...目的:评估不同阶段结肠腺癌患者中CDC样激酶2 (CLK2)基因的表达,以及CLK2在结肠腺癌细胞中的表达变化和CLK2所介导的细胞功能。方法:在TIMER数据库和GEPIA数据库分析CLK2在健康与不同肿瘤组织中的表达水平,找到差异性表达显著的肿瘤类型。分析TCGA数据库中各个阶段结肠腺癌患者组织中CLK2的表达水平变化。使用RT-qPCR和western blot方法检测了CLK2基因在肿瘤组织和细胞系中的表达变化。在敲低CLK2的条件下,使用western blot方法检测细胞增殖标志物和细胞周期标志物的表达。最后,使用STRING工具分析与CLK2基因相关性较强的多个关键基因。结果:结肠腺癌组织中的CLK2基因表达显著升高(p Objective: To assess CDC-like kinase 2 (CLK2) gene expression in patients with different stages of colon adenocarcinoma, as well as changes in CLK2 expression in colon adenocarcinoma cells and cellular functions mediated by CLK2. Methods: Screening for CLK2 differentially expressed tumor types in the TIMER database and the GEPIA database. Detection of CLK2 gene changes with colon adenocarcinoma tumor stage in the TCGA database. Changes in CLK2 gene expression in tumor tissues and cell lines were detected using RT-qPCR and western blot methods. The expressions of cell proliferation markers and cell cycle markers were detected using western blot method under the condition of knocking down CLK2. Finally, multiple key genes with strong correlation with CLK2 gene were analyzed using STRING tool. Results: CLK2 gene expression was significantly elevated in colon adenocarcinoma tissues (p < 0.001), and the high expression of CLK2 gene was significantly correlated with the clinical features of colon adenocarcinoma (pathological stage, T stage) (p < 0.05). CLK2 gene expression is elevated in colon adenocarcinoma cells, and knockdown of CLK2 expression can effectively inhibit cell proliferation and cell cycle. Finally, CLK3, CLK1, SRSF12 and other genes are potential binding targets of CLK2 gene. Conclusion: The expression level of the CLK2 gene is significantly correlated with the progression of colon adenocarcinoma, and decreasing the expression of the CLK2 gene is expected to be a new approach to the treatment of patients with colon adenocarcinoma.展开更多
文摘目的:评估不同阶段结肠腺癌患者中CDC样激酶2 (CLK2)基因的表达,以及CLK2在结肠腺癌细胞中的表达变化和CLK2所介导的细胞功能。方法:在TIMER数据库和GEPIA数据库分析CLK2在健康与不同肿瘤组织中的表达水平,找到差异性表达显著的肿瘤类型。分析TCGA数据库中各个阶段结肠腺癌患者组织中CLK2的表达水平变化。使用RT-qPCR和western blot方法检测了CLK2基因在肿瘤组织和细胞系中的表达变化。在敲低CLK2的条件下,使用western blot方法检测细胞增殖标志物和细胞周期标志物的表达。最后,使用STRING工具分析与CLK2基因相关性较强的多个关键基因。结果:结肠腺癌组织中的CLK2基因表达显著升高(p Objective: To assess CDC-like kinase 2 (CLK2) gene expression in patients with different stages of colon adenocarcinoma, as well as changes in CLK2 expression in colon adenocarcinoma cells and cellular functions mediated by CLK2. Methods: Screening for CLK2 differentially expressed tumor types in the TIMER database and the GEPIA database. Detection of CLK2 gene changes with colon adenocarcinoma tumor stage in the TCGA database. Changes in CLK2 gene expression in tumor tissues and cell lines were detected using RT-qPCR and western blot methods. The expressions of cell proliferation markers and cell cycle markers were detected using western blot method under the condition of knocking down CLK2. Finally, multiple key genes with strong correlation with CLK2 gene were analyzed using STRING tool. Results: CLK2 gene expression was significantly elevated in colon adenocarcinoma tissues (p < 0.001), and the high expression of CLK2 gene was significantly correlated with the clinical features of colon adenocarcinoma (pathological stage, T stage) (p < 0.05). CLK2 gene expression is elevated in colon adenocarcinoma cells, and knockdown of CLK2 expression can effectively inhibit cell proliferation and cell cycle. Finally, CLK3, CLK1, SRSF12 and other genes are potential binding targets of CLK2 gene. Conclusion: The expression level of the CLK2 gene is significantly correlated with the progression of colon adenocarcinoma, and decreasing the expression of the CLK2 gene is expected to be a new approach to the treatment of patients with colon adenocarcinoma.
文摘目的旨在构建结肠腺癌(colonic adenocarcinoma,COAD)中铜死亡相关基因(Cuproptosis-related genes,CRGs)的预后模型,探讨CRGs在COAD中的表达与预后的关系。方法从TCGA数据库下载了正常和COAD患者的转录组和临床数据,并从GEO数据库下载数据集GSE39582作为外部验证集。基于铜死亡相关基因表达量,对TCGA数据进行共识聚类以确定新的肿瘤亚型。采用单因素Cox回归分析、LASSO回归分析筛选预后基因并构建预后风险模型。并通过Kaplan-Meier生存分析、受试者工作特征(receiver operating characteristic,ROC)曲线下的面积(area under the curve,AUC)和一致性指数(concordance index,C-index)对模型进行评价。用GSE44001数据集对模型进行外部验证。基于风险评分及临床因素构建列线图,预测患者的生存率。并进行免疫浸润、GSEA分析、免疫检查点、药敏分析、肿瘤微环境和干细胞指数的研究,系统地阐述COAD中CRGs和免疫的相关性。结果21个CRGs在COAD组织及正常组织中差异表达。单变量Cox回归及Lasso回归分析确定了五个预后相关基因(SDHB,CCS,DLAT,FDX1,CP)并构建模型。Kaplan-Meier分析表明,高风险患者OS显著低于低风险患者(P<0.01),并在GEO队列中取得了验证(P<0.05)。风险模型的AUC值为0.734,表明预后值有统计学意义,模型具有一定的准确性。一致性分析和ROC结果表明,列线图模型有较好的预后预测准确性。这5个CRGs主要参与线粒体、免疫应答和代谢途径,是COAD预后的独立因素,并与免疫细胞浸润、免疫检查点以及肿瘤微环境和干细胞指数密切相关。结论构建了5个铜死亡相关基因的结肠腺癌预后模型,有可能作为未来临床实践中有效的预后标志物。
