基于TCGA数据挖掘和分子对接技术探讨CKS1B高表达在肺腺癌中的意义及芒柄花素的干预机制

Discussion on Significance of High Expression of CKS1B in Lung Adenocarcinoma and Intervention Mechanism of Formononetin Based on TCGA Data Mining and Molecular Docking Techniques

目的通过TCGA数据挖掘和分子对接技术探讨CKS1B高表达在肺腺癌(LUAD)中的意义及芒柄花素的干预机制。方法从TCGA公共数据库获得肺腺癌患者相关数据集,采用基因表达谱交互分析(GEPIA)在线数据库,分析CKS1B基因表达与LUAD发病之间的关系,同时分析CKS1B基因差异表达的生存曲线,寻找CKS1B基因表达与LUAD患者预后的相关性;采用单因素COX回归分析和多因素COX回归分析CKS1B基因表达与LUAD患者性别,年龄,分期,T分期,N分期等临床相关性,判断CKS1B是否为LUAD独立的临床预后因子;采用GSEA进行基因富集分析,初步判断CKS1B影响的相关通路和作用机制;采用分子对接方法对芒柄花素与CKS1B进行对接,预测其结合模式和亲合力,辅助判断芒柄花素的调控机制。结果 TCGA数据库共获得肿瘤样本535例,癌旁组织59例。与癌旁组织比较,LUAD组织中CKS1B基因表达水平显著升高;CKS1B基因低表达的LUAD患者生存率明显高于CKS1B基因高表达的LUAD患者;男性LUAD患者CKS1B基因表达明显高于女性患者;LUADⅡ期患者CKS1B基因表达显著高于Ⅰ期,Ⅳ期显著高于其他三期;LUAD T2期患者CKS1B基因表达显著高于T1期;LUAD N1期患者CKS1B基因表达显著高于N0期。单因素COX回归分析结果显示,临床分期,T分期,M分期,CKS1B基因表达与总的生存期密切相关;多因素COX回归分析显示,临床分期和CKS1B基因表达可以做为独立的临床预后因子;GSEA富集分析结果显示,CKS1B与糖和核糖代谢,氧化磷酸化,平滑肌损伤,哮喘,p53等信号通路密切相关。分子对接结果显示,芒柄花素与CKS1B存在结合靶点,最低结合能为-5.7 kcal/mol。结论 CKS1B基因是LUAD重要的临床预后因子,芒柄花素可能通过调控CKS1B发挥抗LUAD的作用。

Objective To investigated the significance of high expression of CKS1 B in lung adenocarcinoma(LUAD) and the intervention mechanism of Formononetin by TCGA data mining and molecular docking techniques. Methods Data sets of LUAD patients were obtained from the TCGA public database. Gene expression profile interaction analysis(GEPIA) online database was used to analyze the relationship between the expression of CKS1 B gene and the diagnosis of LUAD, and the survival curve of differential expression of CKS1 B gene was analyzed to find out the correlation between the expression of CKS1 B gene and the prognosis of LUAD patients. Univariate COX regression analysis and multivariate COX regression analysis were used to analyze the clinical correlation between CKS1 B gene expression and LUAD patients’ gender, age, stage, T stage, N stage and so on to determine whether CKS1 B was an independent clinical prognostic factor for LUAD. GSEA was used for gene enrichment analysis to preliminarily determine the related pathway and mechanism of CKS1 B influence. The molecular docking method was used to dock formononetin and CKS1 B to predict their binding mode and affinity and help to judge the regulatory mechanism of formononetin. Results A total of 535 tumor samples and 59 para-carcinoma tissues were obtained from the TCGA database. Compared with para-carcinoma tissues, the expression level of CKS1 B gene in LUAD tissues was significantly increased. The survival rate of LUAD patients with low expression of CKS1 B gene was significantly higher than that of LUAD patients with high expression of CKS1 B gene. The expression of CKS1 B gene in male LUAD patients was significantly higher than that in female patients. The expression of CKS1 B gene in LUAD Ⅱ stage patients was significantly higher than that ofⅠ stage patients. That of Ⅳ stage patients was significantly higher than that of other three stages patients. The expression of CKS1 B gene in LUAD T2 stage patients was significantly higher than that in T1 stage patients. The expression of CKS1 B gene in LUAD stage N1 patients was significantly higher than that in Stage N0 patients. Univariate COX regression analysis showed that clinical stage, T stage, M stage and CKS1 B gene expression were closely related to the overall survival. GSEA results showed that CKS1 B was closely related to sugar nucleotide sugar metabolism, oxidative phosphorylation, smooth muscle damage, asthma, p53 and other signaling pathways. Multivariate COX regression analysis showed that clinical stage and CKS1 B gene expression could be independent prognostic factors. Molecular docking results showed that Formononetin and CKS1 B had multiple binding targets. Conclusion CKS1 B gene is an important clinical prognostic factor for LUAD, and Formononetin may play an anti-LUAD role by regulating CKS1 B.