子宫内膜癌组织CKS1B mRNA水平表达与临床病理学特征及预后的相关性研究

Relationship of CKS1B mRNA Expression in Endometrial Cancer Tissues with Its Clinicopathological Features and Prognosis

目的研究细胞周期依赖蛋白酶调节亚基1B(cyclin-dependent kinases regulatory subunit 1B,CKS1B)与子宫内膜癌(endometrial carcinoma,EC)患者临床病理特征及预后的相关性。方法下载癌症基因组图谱(the Cancer Genome Atlas,TCGA)数据库和基因型-组织表达(genotype-tissue expression,GTEx)数据库中CKS1B的表达谱数据和临床数据,分析CKS1B在子宫内膜癌中的表达及与临床病理特征和预后的关系。通过UALCAN数据库验证CKS1B蛋白表达水平;Logistics回归分析CKS1B表达与临床病理参数的关系;COX回归分析和Kaplan-Meier分析临床病理参数与子宫内膜癌预后的相关性;分析TCGA数据库中CKS1B的共表达基因并富集分析。实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)检测CKS1B mRNA在Ishikawa和HEC-1-A细胞系中的表达。蛋白质免疫印迹(Western Bolt,WB)检测CKS1B蛋白在子宫内膜癌组织和癌旁组织的表达。结果CKS1B mRNA和蛋白在子宫内膜癌中明显高于正常子宫内膜,差异具有统计学意义(P<0.05)。CKS1B mRNA表达水平与患者FIGO分期(F=42.994)、组织学分级(F=70.350)、组织学类型(F=87.341)和年龄(F=40.097)密切相关(均P<0.05)。Kaplan-Meier法显示CKS1B m RNA高表达组患者总生存率更低(Log-rankχ^(2)=1.175,P<0.01)。多因素COX分析显示CKS1B表达水平(HR=1.856,95%CI:1.154~2.985)、FIGO分期(HR=3.065,95%CI:1.906~4.926)为子宫内膜癌预后的独立风险因素(P<0.05)。GO分析显示CKS1B主要参与细胞核分裂、染色体分离。KEGG分析显示主要在细胞周期、剪接体及DNA复制通路富集。进一步验证与人子宫内膜上皮细胞比较CKS1B mRNA在Ishikawa和HEC-1-A细胞中高表达(F=44.560,P<0.001),CKS1B蛋白在子宫内膜癌组织中高表达(t=14.900,P<0.001)。结论CKS1B在子宫内膜癌中表达上调且与患者的临床病理因素及预后密切相关,可能通过细胞周期调控参与子宫内膜癌发生、发展,有望成为子宫内膜癌诊断和预测预后的新指标。

Objective To investigate the expression of CKS1B in endometrial carcinoma(EC)and its relationship with clinicopathological features and prognosis.Methods The expression profile data and clinical data of CKS1B from the TCGA and GTEx databases were downloaded to investigate the expression of CKS1B in EC and its relationship with clinicopathological features.The expression of CKS1B at the protein level was verified using the UALCAN database.The relationship between CKS1B expression and clinicopathological parameters was analyzed by Logistic regression.The r program perform enrichment analysis on CKS1B co-expressed genes in the TCGA database.Finally,CKS1B mRNA expression was discovered in the cell lines Ishikawa and HEC-1-A by quantitative real-time PCR(qRT-PCR).CKS1B protein expression was detected in EC tissues and adjacent tissues by Western Bolt(WB).Results CKS1B mRNA and protein were remarkably higher in EC tissues than in normal endometrium,and the differences were statistically significant(P<0.05).The level of CKS1B mRNA expression was strongly correlated with FIGO stage(F=42.994),histological grade(F=70.350),histological type(F=87.341)and age(F=40.097)(all P<0.05).The results of the Kaplan-Meier method showed that patients with high CKS1B mRNA expression had a lower overall survival rate(Log-rankχ^(2)=1.175,P<0.01).Multifactorial COX analysis showed that FIGO stage(HR=3.065,95%CI:1.906~4.926)and CKS1B expression(HR=1.856,95%CI:1.154~2.985)were independent risk factors affecting the prognosis of patients with EC(P<0.05).GO analysis showed that CKS1B was mainly involved in nuclear division and chromosome separation.KEGG analysis showed it was mainly enriched in the cell cycle,spliceosome and DNA replication.Further verification showed that CKS1B mRNA was highly expressed in Ishikawa and HEC-1-A cell lines(F=44.560,P<0.001),CKS1B protein was highly expressed in EC tissues(t=14.900,P<0.001).Conclusion CKS1B is upregulated in EC and is linked to clinicopathological variables in the patients.It may play a role in the development of EC by regulating the cell cycle,and it is expected to be a new marker for the diagnosis and prognosis of EC.