摘要
目的探究SMAD4下调在胰腺癌侵袭转移中的分子机制。方法通过微阵列分析研究胰腺癌细胞株PANC-1中经SMAD4-siRNA处理后相关基因的变化。使用Limma软件包鉴定差异表达基因(differentially expressed genes,DEGs),进行基因本体论(Gene Ontology,GO)生物过程富集分析和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集分析。通过STRING数据库获得DEGs编码蛋白质之间的互相作用关系,运用Cytoscape软件构建蛋白质蛋白质相互作用(protein-protein interaction,PPI)网络,并通过Cytoscape的插件MCODE进行PPI子网络分析。通过实时荧光定量PCR(qRT-PCR)验证一些关键基因的表达情况。结果通过qRT-PCR和Western印迹法验证PANC-1细胞株中的SMAD4 mRNA水平、蛋白水平被成功下调。经差异分析,共鉴定出402个DEGs,其中有168个上调基因,234个下调基因,GO功能和KEGG信号通路富集分析显示,表达上调的基因主要富集在大分子复合物的亚基组织、大分子复合物的组装、细胞黏附、生物黏附等生物学过程,以及系统性红斑狼疮信号通路中;而表达下调的基因主要富集在免疫反应、防御反应、细胞增殖调控、对损伤的反应等生物学过程,以及细胞因子细胞因子受体相互作用信号通路、RIG-I样受体信号通路、趋化因子信号通路、Toll样受体信号通路、自然杀伤细胞介导的细胞毒性和胞质DNA感应信号通路。PPI网络中排名前五的节点基因为ISG15、MX1、IFIT1、STAT1、IRF7和IRF1。IFIT2和IFIH1为PPI子网络核心模块的枢纽蛋白。上述基因主要集中在Ⅰ型干扰素(interferon,IFN)信号通路上。结论SMAD4下调可能通过阻止Ⅰ型IFN介导的免疫监测,在促进胰腺癌的转移中发挥作用。
Objective To explore the molecular mechanism of SMAD4 in the invasion and metastasis of pancreatic cancer.Methods The changes of related genes in pancreatic cancer PANC-1 cells after SMAD4-siRNA treatment were analyzed by microarray.The main biological processes and signal pathways of differentially expressed genes(DEGs),gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment were analyzed by Limma software package.The interaction between DEGs coded protein was obtained through STRING database,and the Protein-Protein Interaction(PPI)network was constructed by Cytoscape software,and the PPI sub-network was analyzed through the plug-in MCODE of Cytoscape.The expression of some key genes was verified by qRT-PCR.Results A total of 402 DEGs were identified,including 168 up-regulated genes and 234 down-regulated genes.The analysis of GO function and KEGG signal pathway showed that the up-regulated genes were mainly enriched in biological processes such as subunit organization of macromolecular complex,assembly of macromolecular complex,cell adhesion and biological adhesion,and the signal pathway of systemic lupus erythematosus.The down-regulated genes were mainly concentrated in biological processes such as immune response,defense response,cell proliferation regulation and response to injury,as well as cytokine-cytokine receptor interaction signal pathway,RIG-I-like receptor signal pathway,chemokine signal pathway,Toll-like receptor signal pathway,natural killer cell-mediated cytotoxicity and cytoplasmic DNA sensing signal pathway.The top five node genes in PPI network were ISG15,MX1,IFIT1,STAT1,IRF7 and IRF1.IFIT2 and IFIH1 were hub proteins in the core module of PPI subnetwork.The above genes were mainly concentrated in the signaling pathway of typeⅠinterferon(IFN).Conclusion SMAD4 may play a role in promoting the metastasis of pancreatic cancer through preventing immune monitoring mediated by typeⅠIFN.
作者
贺雅芝
王锋
王阳
宋亦然
张润
余畋余
童依丽
HE Yazhi;WANG Feng;WANG Yang;SONG Yiran;ZHANG Run;YU Tianyu;TONG Yili(Anhui University of Science and Technology,Huainan 232001,Anhui Province,China;Department of Gastroenterology,Huadong Hospital Affiliated to Fudan University,Shanghai 200040,China;Department of Gastroenterology,Shanghai Tenth People s Hospital,School of Medicine,Tongji University,Shanghai 200072,China;Institute for Bioengineering and Nanotechnology,The University of Queensland,Queensland 4072,Australia;Department of General Practice,Huadong Hospital Affiliated to Fudan University,Shanghai 200040,China)
出处
《同济大学学报（医学版）》
2023年第3期326-335,共10页
Journal of Tongji University(Medical Science)
基金
国家自然科学基金(81972287)
上海市自然科学基金(19ZR1447600)。
关键词
胰腺癌
干扰素
微阵列
SMAD4
转移
pancreatic cancer
interferon
microarray
SMAD4
metastasis
