生物信息学筛选骨肉瘤关键基因ITGA3及差异表达验证  

Bioinformatic screening of key gene ITGA3 in osteosarcoma and differential expression vali⁃dation

作　　者：徐伟华 朱必康 瞿洋洋 谭曼利 罗世兴[3] Xu Weihua;Zhu Bikang;Qu Yangyang;Tan Manli;Luo Shixing(Collaborative Innovation Centre of Regenerative Medicine and Medical BioResource Development and Application Co-constructed by the Province and Ministry,Guangxi Medical University,Nanning 530021,China;Guangxi Engineering Center in Biomedical Materials for Tissue and Organ Regeneration,Guangxi Medical University,Nanning 530021,China;Beihai People's Hospital,Beihai 536000,China)

机构地区：[1]广西医科大学再生医学与医用生物资源开发应用省部共建协同创新中心,南宁530021 [2]广西医科大学广西组织器官修复医用生物材料工程技术研究中心,南宁530021 [3]北海市人民医院,北海536000

出　　处：《广西医科大学学报》2022年第5期769-774,共6页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81760485);广西科技基地和人才专项资助项目(No.桂科AD19254003);广西医科大学青年科学基金资助项目(No.GXMUYSF202107)。

摘　　要：目的:生物信息学分析筛选出骨肉瘤关键基因,探讨骨肉瘤潜在分子标记物。方法:从GEO数据库中下载GSE14359、GSE16088、GSE160913个数据库集,利用R语言“SVA”软件包进行合并;差异分析利用R语言“limma”软件包进行分析,并对差异基因运用R语言“clusterProfiler”进行GO和KEGG富集分析。随后,对最显著富集的KEGG通路基因进行筛选出最关键的Top5基因并进行生存分析,筛选对预后具有影响意义的关键基因,然后运用实时荧光定量聚合酶链式反应(qRT-PCR)验证最关键基因的差异性表达。结果:通过数据合并共得到12547个基因的数据集,分析出723个表达基因有差异;差异基因的KEGG富集最显著通路是PI3K-Akt信号通路,并且该通路的排名前5的关键基因为ITGA3、ITGA4、ITGAV、COL1A1、COL1A2。排名前5的关键基因中,与ITGA3低表达组比较,ITGA3高表达组生存预后较好(P<0.05),在143B、MNNG、U2OS 3个骨肉瘤细胞系中ITGA3相对表达量低于正常成骨细胞系hFOB1.19(P<0.01)。结论:ITGA3在骨肉瘤中呈现出低表达,这可能成为预测骨肉瘤患者预后的潜在生物标记物。Objective:To screen the key genes of osteosarcoma by bioinformatics analysis and explore potential molecular markers for osteosarcoma.Methods:Three data sets GSE14359,GSE16088,and GSE16091 were downloaded from GEO database and merged using the R language“SVA”package;the differential analysis was performed using the R language“limma”package,and the GO and KEGG enrichment analysis was performed using the R language“cluster Profiler”for the differential genes.Subsequently,the most significantly enriched KEGG pathway genes were screened for the five most critical genes and survival analysis were performed to screen the critical genes with prognostic impact,the differential expression of which was validated using realtime quantitative PCR(RT-qPCR).Results:A total dataset of 12,547 genes was obtained by data merging,among which 723 were differentially expressed genes.The most significantly enriched KEGG pathway for differential genes was the PI3K-Akt signaling pathway,and the top 5 key genes of this pathway were ITGA3,ITGA4,ITGAV,COL1A1,and COL1A2.Among the top 5 key genes,ITGA3 high expression group had a better survival prognosis compared with ITGA3 low expression group(P<0.05),and the relative expression of ITGA3 in the 3 osteosarcoma cell lines 143B,MNNG,and U2OS was lower than that in the normal osteoblast cell line hFOB1.19(P<0.01).Conclusion:ITGA3 showed low expression in osteosarcoma,which may be a potential biomarker to predict the prognosis of osteosarcoma patients.

关 键 词：骨肉瘤 ITGA3 GEO 

分 类 号：R738.1[医药卫生—肿瘤]