CBX8抑制前列腺癌细胞侵袭的机制研究  

Deciphering the suppressive effects of CBX8 on prostate cancer cell invasion

作　　者：杨万里[1] 宋娟 李兵[1] 劳一敏 YANG Wanli;SONG Juan;LI Bing;LAO Yimin(Department of Biochemistry and Molecular Cell Biology,Shanghai Jiao Tong University College of Basic Medical Sciences,Shanghai 200025,China)

机构地区：[1]上海交通大学基础医学院生物化学与分子细胞生物学系,上海200025

出　　处：《上海交通大学学报（医学版）》2023年第12期1507-1519,共13页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家自然科学基金(81802773)。

摘　　要：目的·探究染色质组织调节框同源蛋白8(chromobox protein homolog 8,CBX8)在前列腺癌中的生物学功能,并通过转录组及表观修饰分析揭示CBX8在前列腺癌转移中的作用机制。方法·利用cBioPortal数据库对癌症基因组图谱(The Cancer Genome Atlas,TCGA)中前列腺腺癌(prostate adenocarcinoma,PRAD)患者样本数据集进行CBX家族蛋白mRNA表达分析。采用短发夹RNA技术敲低DU145前列腺癌细胞系中的CBX8,通过CCK-8和Transwell实验检测细胞增殖和侵袭水平的变化。使用RNA转录组测序(RNA-seq)分析敲低CBX8后影响的差异表达基因。对这些差异表达基因进行基因集富集分析(Gene Set Enrichment Analysis,GSEA)、基因本体论(Gene Ontology,GO)功能分析以及京都基因和基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)信号通路富集分析。通过染色质免疫沉淀测序(ChIPseq)观察和测定敲低CBX8后基因组H3K27me3甲基化水平的变化。结果·根据对TCGA-PRAD患者样本数据的分析,发现CBX8 mRNA在前列腺癌中高表达。在前列腺癌细胞系DU145中敲低CBX8后,细胞的增殖能力没有显著变化(P>0.05),但其侵袭能力却显著提高(P<0.05)。RNA-seq分析显示CBX8敲低导致750个基因表达上调,951个基因表达下调;其中,与多种肿瘤转移有关的支链氨基酸转氨酶1(branched-chain-amino-acid aminotransferase 1,BCAT1)在敲除CBX8后表达明显上升。GSEA显示表达水平受影响的基因与多梳蛋白复合体1(polycomb repressive complex 1,PRC1)的功能有关。同时,通过GO和KEGG信号通路富集分析发现受影响的生物过程包括转运RNA(transfer RNA,tRNA)氨酰化、DNA复制、氨酰基-tRNA连接酶活性变化以及钙黏蛋白的结合等;特别是在GO功能分析的细胞组分方面富集了与肿瘤转移有关的细胞-基底连接相关基因。利用ChIP-seq对表观修饰的研究显示,在敲低CBX8后全基因组的H3K27me3水平有所下降;并鉴定了97个位于CBX8敲低后转�Objective·To elucidate the regulatory mechanisms of the chromobox protein homolog 8(CBX8)in prostate cancer metastasis from transcriptome and epigenetic modification perspectives.Methods·The correlation between the expression of CBX proteins and prostate adenocarcinoma(PRAD)in The Cancer Genome Atlas(TCGA)was examined through an analysis based on cBioPortal database.A stable CBX8 knockdown DU145 prostate cancer cell line was established via short hairpin RNA(shRNA)transfection.Subsequently,the proliferation and invasion of the CBX8 knockdown cells were analyzed by CCK-8 assay and Transwell assay,respectively.Transcriptome changes of the CBX8 knockdown cells were investigated through RNA sequencing(RNA-seq)coupled with Gene Set Enrichment Analysis(GSEA).To further evaluate the functional implications of these transcriptomic alterations,Gene Ontology(GO)for functional analysis was deployed.Moreover,to identify potentially affected signalling pathways,the Kyoto Encyclopedia of Genes and Genomes(KEGG)was utilized for pathway enrichment analysis.Lastly,the levels of H3K27me3,a key histone modification associated with CBX8,in the knockdown cells were determined by chromatin immunoprecipitation sequencing(ChIP-seq).Results·Bioinformatic analysis with cBioPortal database,based on TCGA-PRAD cohorts,unveiled a high CBX8 mRNA expression in PRAD.Knockdown of CBX8 did not significantly affect the proliferation of DU145 cells(P>0.05),but caused a a significant increase in their invasiveness(P<0.05).The RNA-seq analysis revealed that CBX8 knockdown led to the upregulation of 750 genes and the downregulation of 951 genes.Notably,branched-chain-amino-acid aminotransferase 1(BCAT1),a gene implicated in the metastasis of various types of cancers,showed a significant increase in expression following CBX8 knockdown.GSEA showed that the expression levels were of the affected genes were related to the functions of the polycomb repressive complex 1(PRC1).A further investigation using GO and KEGG analyses identified several enriched p

关 键 词：染色质组织调节框同源蛋白8 前列腺癌 细胞增殖 肿瘤侵袭 

分 类 号：R34[医药卫生—基础医学]