基于IPA探析COPB2基因对人骨肉瘤细胞的分子调控机制  

Molecular Regulation Mechanism of COPB2 Gene on Human Osteosarcoma Cell Based on Ingenuity Pathway Analysis

作　　者：崔云鹏[1] 潘元星 林云飞[1] 米川[1] 王冰[1] 施学东[1] CUI Yunpeng;PAN Yuanxing;LIN Yunfei;MI Chuan;WANG Bing;SHI Xuedong(Department of Orthopaedics,Peking University First Hospital,Beijing 100034,China)

机构地区：[1]北京大学第一医院骨科,北京100034

出　　处：《肿瘤防治研究》2020年第4期251-255,共5页Cancer Research on Prevention and Treatment

摘　　要：目的基于IPA(ingenuity pathway analysis)的生物信息学分析,探析COPB2对人骨肉瘤细胞的分子调控机制。方法RT-qPCR检测COPB2在人骨肉瘤细胞系U-2OS、MG-63、HOS和Saos-2中的表达水平。以U-2OS为研究对象,分为沉默组(KD)和对照组(NC)。绿色荧光蛋白标记的慢病毒感染U-2OS细胞,沉默组含COPB2沉默序列。人基因表达谱芯片检测COPB2沉默后U-2OS细胞基因表达谱变化差异,将检测信息输入IPA软件进行生物信息学分析,包括经典通路、上游调控、疾病功能、调控效应和互作网络等分析。结果COPB2在四种人骨肉瘤细胞系中表达丰度均高。人基因表达谱芯片显示,KD组与NC组相比,1393个基因表达出现显著差异,其中上调基因数831个、下调基因数562个。IPA经典通路分析显示COPB2敲除后,TSP1抑制血管形成通路、急性期反应通路被显著激活,细胞周期蛋白和细胞周期调控通路显著抑制。疾病功能和互作网路分析显示COPB2在骨肉瘤中发挥重要作用,主要影响细胞瘤变、肿瘤细胞侵袭和失巢凋亡等功能。结论COPB2在人骨肉瘤细胞系中高表达,可能通过细胞周期蛋白和细胞周期调控通路参与骨肉瘤的发生发展。COPB2与骨肉瘤细胞瘤变、侵袭力和失巢凋亡相关。Objective To explore the possible molecular regulation mechanism of COPB2 on human osteosarcoma cells based on the bioinformatics analysis of ingenuity pathway analysis(IPA).Methods RTqPCR was used to detect the expression levels of COPB2 in human osteosarcoma cell lines U-2OS,MG-63,HOS and Saos-2.U-2OS was used as the research object and divided into silence group(KD)and control group(NC).U-2OS cells were infected by green fluorescent protein-labeled lentivirus and the KD group contained COPB2 silenced sequence.Human gene expression profiling microarray was used to detect the difference in gene expression profile of U-2OS cells after COPB2 silencing.The detection information was input into the IPA software for bioinformatics analysis,including classical pathway analysis,upstream regulation analysis,disease function analysis,regulatory effect analysis and interaction network analysis.Results COPB2 had a high expressive abundance in four human osteosarcoma cell lines.There were significant differences in the expression of 1393 genes between KD group and NC group;831 genes were upregulated and 562 genes were down-regulated.After COPB2 knockdown,the inhibition of angiogenesis by TSP1 pathway and acute phase response pathway were significantly activated,while cyclins and cell cycle regulation pathway were significantly inhibited.COPB2 mainly affected neoplasia,invasion and anoikis of osteosarcoma cells.Conclusion COPB2 is highly expressed in human osteosarcoma cell lines,and might be involved in the development of osteosarcoma through cyclins and cell cycle regulation pathway.In addition,COPB2 is associated with neoplasia,invasion and anoikis of osteosarcoma cells.

关 键 词：外被体包被蛋白复合体β2亚基 骨肉瘤 生物信息学 

分 类 号：R738.1[医药卫生—肿瘤]