南方医科大学学报

南方医科大学学报 ›› 2020, Vol. 40 ›› Issue (05): 683-692.doi: 10.12122/j.issn.1673-4254.2020.05.11

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BUB1基因在胃癌组织中高表达： 基于Oncomine数据库及生物信息学方法

李小燕，何 杰，余 觅，张 维，孙 建   

  • 出版日期:2020-05-20 发布日期:2020-05-20
  • 基金资助:

BUB1 gene is highly expressed in gastric cancer: analysis based on Oncomine database and bioinformatics

  

  • Online:2020-05-20 Published:2020-05-20

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摘要： 目的 探讨细胞周期有丝分裂纺锤体检测点丝/苏氨酸激酶（BUB1）基因在胃癌（STAD）中的表达及临床价值。方法 分别运用Oncomine、GEPIA、BioGPS和Kaplan-Meier Plotter等数据库分析BUB1基因在胃癌组织和正常胃组织的表达差异，并且分析BUB1表达水平与胃癌患者生存预后之间的关系；利用癌症细胞系全科百科全书（CCLE）分析BUB1分别在胃癌组织T细胞和B细胞中的表达，利用String数据库绘制BUB1相关蛋白网络图及基因本体（GO）功能注释，并对京都基因和基因组百科全书（KEGG）相关通路进行分析。采用肿瘤免疫评估资源（TIMER）数据库分析BUB1在免疫浸润物中的表达及对胃癌患者生存预后的影响。为进一步验证Oncomine数据库基因芯片的结果，我们选择成都医学院第一附属医院2018年3月~2019年7月收治的胃癌患者30例，经外科手术获得胃腺癌组织30份，同时获取的癌旁正常组织30份作为对照组，通过PCR和免疫组化技术对Oncomine数据库中BUB1基因表达的数据进行验证。结果 Oncomine、GEPIA、BioGPS分析显示：与正常胃组织相比，BUB1在胃癌组织中呈现高表达（P<0.05）。通过Kaplan-Meier生存分析发现，BUB1基因高表达的胃癌患者无进展生存期（HR=0.52，95%CI=0.41~0.67，P<0.05）、总生存期（HR=0.67，95%CI=0.55~0.82，P<0.05）均有所延长。通过string数据收集到与BUB1相关 蛋白主要富集于13类细胞学组分、4类分子功能和12类生物学过程，并参与了4条信号通路。TIMER数据库分析显示在免疫微环境中高表达BUB1 mRNA的CD4+T细胞和巨噬细胞对胃癌患者5年生存预后较好。实时定量PCR结果提示胃癌组织样本 BUB1的表达水平（4.345±0.421）明显高于配对的癌旁正常胃粘膜组织（1.583±0.122）（t=34.63，P<0.05）；免疫组化结果提示胃癌组织BUB1染色成阳性。结论 胃癌组织中BUB1基因呈现高表达，BUB1可能具有减少肿瘤免疫抑制作用可协助评估胃癌患者的预后情况。

Abstract: Objective To investigate the expression of BUB1 gene in gastric cancer. Methods Oncomine, GEPIA, BioGPS and Kaplan-Meier Plotter databases were used to analyze the difference of BUB1 gene expression between gastric cancer tissue and normal gastric tissue. The association of BUB1 expression level with the prognosis of gastric cancer patients was also analyzed. The Cancer Cell Line Encyclopedia (CCLE) was explored to analyze the expression of BUB1 in T cells and B cells in gastric cancer patients, and the String database was used to generate the network map of BUB1- related proteins and functional annotation of gene ontology (GO). The related pathways of KEGG were analyzed. Tumor immune assessment resource (TIMER) database was used to analyze the expression of BUB1 in immune infiltration and its effect on prognosis of gastric cancer patients. To further verify the results of gene chip analysis in Oncomine database, we collected 30 pairs of surgical specimens of gastric adenocarcinoma and adjacent tissues from patients admitted to the First Affiliated Hospital of Chengdu Medical College from March, 2018 to July, 2019. The results of BUB1 gene expression in Oncomine database were verified by PCR and immunohistochemistry. Results Oncomine, GEPIA and BioGPS analyses showed that BUB1 was highly expressed in gastric cancer compared with normal gastric tissue. Kaplan-Meier survival analysis showed that the progression-free survival time (HR=0.52, 95%CI: 0.41-0.67, P<0.05) and the overall survival time (HR=0.67, 95%CI: 0.55-0.82, P<0.05) were prolonged in gastric cancer patients with a high expression of BUB1. Through String data collection, BUB1-related proteins were mainly enriched in 13 cellular components, 4 molecular functions and 12 biological processes, involving 4 signal pathways. TIMER database analysis showed that CD4 + T cells and macrophages with high expressions of BUB1 mRNA in the immune microenvironment were associated with a favorable 5-year survival outcome of patients with gastric cancer. In the surgical specimens, real-time quantitative PCR showed that the expression level of BUB1 mRNA was significantly higher in gastric cancer tissues than in the adjacent gastric mucosa tissues, and immunohistochemical results demonstrated positive BUB1 staining in the gastric cancer tissues. Conclusion BUB1 gene is highly expressed in gastric cancer. BUB1 may reduce tumor immunosuppression and helps to evaluate the prognosis of patients with gastric cancer.