基因芯片技术检测NOB1对骨肉瘤细胞相关基因的调控作用

doi:10.13481/j.1671-587x.20180508

摘要/Abstract

摘要： 目的：探讨应用基因芯片技术筛选出可能受NOB1调控的基因，阐明NOB1对骨肉瘤细胞相关基因表达的调控作用。方法：应用慢病毒介导的短发夹RNA（shRNA）干扰技术建立抑制NOB1 mRNA表达的Lv-shNOB1-U2OS骨肉瘤细胞株，实验分为2组，分别为对照病毒液感染U2OS骨肉瘤细胞（Lv-shCon-U2OS）组和含NOB1干扰质粒病毒感染U2OS骨肉瘤细胞（Lv-shNOB1-U2OS）组。利用mRNA表达谱芯片对Lv-shCon-U2OS组和Lv-shNOB1-U2OS组细胞分别行表达谱分析，并利用生物信息学数据库肿瘤基因（GO）和京都基因与基因组百科全书（KEGG）预测miRNA参与调控的生物学过程、细胞组分、分子功能和信号通路。结果：mRNA表达谱芯片数据显示，NOB1干扰后共有792个基因mRNA表达水平上调，1059个基因mRNA表达水平下调，总共差异变化基因为1851个。GO分析，从细胞位置条目的富集程度来看，差异基因编码产物蛋白主要分布于细胞质膜上，占总差异基因的56.9%，分布于细胞外区域的差异基因编码产物蛋白占总差异基因的39.4%，分布于细胞外空间占总差异基因的20%；从分子功能条目的富集程度来看，差异基因编码产物蛋白主要功能为钙离子结合相关功能（占总差异基因的22%），其次功能为转运子活性（占总差异基因的9.2%），第3位功能为肌动蛋白结合活性，（占总差异基因的8.7%）；从生化过程条目的富集程度来看，差异基因编码产物蛋白主要参与过程为信号转导（占总差异基因的18.7%），其次参与过程为多种细胞器的生成（占总差异基因的15.6%），第3位过程为细胞黏附过程（占总差异基因的10.2%）；KEGG分析，差异分子所编码的蛋白涉及领域包括细胞质膜成分、钙离子结合活性以及信号转导过程。结论：敲除NOB1可以对骨肉瘤细胞相关基因表达产生全方位的影响。

关键词: NOB1基因, 骨肉瘤, 基因表达谱, 短发夹RNA

Abstract: Objective:To screen the genes may be regulated by NOB1 by using gene microarray technique, and to clarify the regulatory effect of NOB1 on the expression of osteosarcoma cell-related genes. Methods:The U2OS cells were treated with lentivirus-mediated RNA interference and to establish the osteosarcoma cells Lv-shNOB1-U2OS. Lv-shCon-U2OS group and Lv-shNOB1-U2OS group were set up. The mRNA expressions of those cells were detected using expression pattern analysis. Gene Oncology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to predict the global functions of NOB1, including biological processes, cellular components, molecular functions, and signaling pathways. Results:After NOB1 interference in the U2OS cells, there were 792 genes with up-regulated mRNA expression level and 1 059 genes with down-regulated mRNA expression level, with a total variation of 1 851 genes. The GO analysis results showed that from the enrichment degree of cell location entries, the differentially encoded product proteins were mainly distributed on the cell membrane as 56.9% of the total difference genes, 39.4% of the totally differential genes distributed in the extracellular region, and 20% of the totally differential genes in the extracellular space; from the enrichment degrees of the molecular function items, the main function of the differentially encoded product proteins was calciu mion binding-related function(22% of the totally differential genes), the second function was transporter activity (9.2% of the totally differenital genes), and the third function was actin binding activity(8.7% of the totally differential genes). In terms of the enrichment of biochemical process entries, the main participation process of differentially encoded product proteins was 18.7% of the totally differential genes of signal transduction, the second involved process was 15.6% of the totally differential genes produced by multiple organelles, and the third process was the cell adhesion process accounted for 10.2% of the totally differential genes. The KEGG analysis results showed that their encoding proteins were involved in plasma membrane, calcium ion binding activity and signal transduction. Conclusion:Knockout of NOB1 can affect the expressions of osteosarcoma cell-related genes in an all-round way.

Key words: NOB1 gene, osteosarcoma, gene expression profile, short haripinRNA

中图分类号:

R738.1

陈炳鹏, 任明, 李容杭, 韩青, 王金成. 基因芯片技术检测NOB1对骨肉瘤细胞相关基因的调控作用[J]. 吉林大学学报(医学版), 2018, 44(05): 929-934.

CHEN Bingpeng, REN Ming, LI Ronghang, HAN Qing, WANG Jincheng. Regulatory effect of NOB1 on osteosarcoma cell-related genes detected by gene microarray technique[J]. Journal of Jilin University(Medicine Edition), 2018, 44(05): 929-934.

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