应用生物信息学筛选结直肠癌Hub基因及验证

Screening and Validation of Hub Gene in Colorectal Cancer by Bioinformatics

目的通过运用生物信息学分析,筛选出与结直肠癌相关的差异表达基因(Differentially Expressed genes,DEGs),并验证其生物学功能。方法云浮市人民医院检验科从基因表达综合数据库(Gene Expression Omnibus,GEO)中下载结直肠癌芯片数据GSE 21815、GSE 31905、GSE 35279资料,应用GEO2R语言进行处理得出结直肠癌和正常结直肠组织之间的差异表达基因,并通过生物信息学工具DAVID、STRING、Cytoscape构建差异表达基因的蛋白互作网络,筛选Hub基因,应用基因本体论(Gene Ontology,GO)、基因百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)分析筛选出的Hub基因的生物功能,并利用MiRDB工具找出可能调控Hub基因的miRNA,并于2017—2022年8月收集30例结直肠癌组织和30例正常结直肠组织样本,通过实时荧光定量PCR(Quantitative Real-time PCR,qPCR)验证。结果经生物学信息分析和蛋白质相互作用网络图分析催产素受体基因、基质金属蛋白酶11基因、间质上皮转化因子基因、基质金属蛋白酶7基因、激肽释放酶8基因、激肽释放酶10基因为结直肠癌组织发生发展的关键Hub基因。结直肠癌组织中基质金属蛋白酶11基因(4.38±1.58)、间质上皮转化因子基因(2.69±0.29)、基质金属蛋白酶7基因(0.88±0.14)、激肽释放酶8基因(11.09±3.90)、激肽释放酶10基因mRNA(7.88±2.20)的表达,显著高于正常结直肠组织组织,差异有统计学意义(t=9.605、25.339、26.376、9.541、3.726,P均<0.001)。结论结直肠癌组织中基质金属蛋白酶11基因、间质上皮转化因子基因、基质金属蛋白酶7基因、激肽释放酶8基因、激肽释放酶10基因异常表达,可能参与结直肠癌发生过程,有望为后续结直肠癌基础研究及临床诊疗提供依据。

Objective Through bioinformatics analysis,the differentially expressed genes(DEGs)associated with colorectal cancer were screened out and their biological functions were verified.Methods The clinical laboratory of Yunfu People's Hospital downloaded colorectal cancer chip data GSE 21815,GSE 31905 and GSE 35279 from Gene Expression Omnibus(GEO)database.The differentially expressed genes between colorectal cancer and normal colorectal tissue were obtained by GEO2R language.The protein interaction network of differentially expressed genes was constructed by bioinformatics tools DAVID,STRING and Cytoscape,and Hub genes were screened.The biologi⁃cal functions of Hub Genes screened were analyzed using gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG).Mirnas that may regulate hub genes were identified using the MiRDB tool,and Quantitative Real-time PCR(qPCR)was used to verify 30 colorectal cancer tissue samples and 30 normal colorectal tissue samples col⁃lected from August 2017 to 2022.Results Oxytocin receptor gene,matrix metalloproteinase-11 gene,mesenchymal epithelial transformation factor gene,matrix metalloproteinase-7 gene,kallikrein-8 gene and kallikrein-10 gene were analyzed as the key Hub genes in the development of colorectal cancer tissue by biological information analysis and protein interaction network diagram.The mRNA expression of matrix metalloproteinase 11 gene(4.38±1.58),stromal epithelial transformation factor gene(2.69±0.29),matrix metalloproteinase 7 gene(0.88±0.14),kallikrein 8 gene(11.09±3.90),and kallikrein 10 gene(7.88±2.20)in colorectal cancer tissue were significantly higher than those in normal colorectal tissue,and the differences were statistically significant(t=9.605,25.339,26.376,9.541,3.726,all P<0.001).Conclusion The abnormal expression of matrix metalloproteinase 11 gene,stromal epithelial transformation factor gene,matrix metalloproteinase 7 gene,kallikrein 8 gene,and kallikrein 10 gene in colorectal cancer tissue may be involved in the development process of colorectal cancer,and is expected to provide a basis for subsequent basic research and clinical diagnosis and treatment of colorectal cancer.