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...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敲低后转录上调基因附近的位点,其中包括BCAT1转录起始位点。结论·CBX8在人前列腺癌中高表达。CBX8具有抑制肿瘤细胞侵袭的功能。其机制可能是CBX8/PRC1复合体结合于BCAT1转录起始位点并抑制BCAT1转录。展开更多
色素框同源蛋白8(chromobox protein homolog 8,CBX8)是PcG家族蛋白PRC1复合体的核心组成部分,在细胞增殖、衰老、维持干细胞自我更新和全能性及肿瘤发生中发挥重要作用.目前研究发现CBX8在多种恶性肿瘤中表达增高,并与肿瘤的进展及预...色素框同源蛋白8(chromobox protein homolog 8,CBX8)是PcG家族蛋白PRC1复合体的核心组成部分,在细胞增殖、衰老、维持干细胞自我更新和全能性及肿瘤发生中发挥重要作用.目前研究发现CBX8在多种恶性肿瘤中表达增高,并与肿瘤的进展及预后密切相关,已成为当前肿瘤领域的研究热点.本文就当前CBX8在肿瘤中的研究作一综述.展开更多
<abstract>Aim: Identification of the rodent counterparts of human and canine epididymal cDNAs HE3, HE4 and Ce8/Ly6G5C by sequence homology and analysis of their expression patterns and regulation level in the ra...<abstract>Aim: Identification of the rodent counterparts of human and canine epididymal cDNAs HE3, HE4 and Ce8/Ly6G5C by sequence homology and analysis of their expression patterns and regulation level in the rat. Methods: 'Electronic screening' of Expressed Sequence Tag (EST) and genomic databases, followed by RT-PCR and Northern blot analysis. Results: Rodent ESTs and genomic sequences homologous to HE3, HE4 and Ce8/Ly6G5C were identified in the public databases and the 'full-length' rat cDNAs cloned. To emphasise their homology to the human and canine genes, they were named Me3/Re3, Me4/Re4 and Re8 for mouse and rat counterparts, respectively. mRNA expression patterns were analysed in rats, including rat HEl and HE5/CD52 counterparts as controls. Re3 and Re8 mRNAs were only found in the rat epididymis, while Re4 showed a broader tissue distribution. Within the epididymis, Re3 and Re4 mRNAs were detected in all regions; Re8, on the other hand, was restricted to the caput. During postnatal development, Re3 and control mRNAs were found from the earliest stages investigated, while Re8 mRNA was observed only from day 24 postnatum, corresponding to the onset of spermatogenesis in the prepubertal testis. Castration and testosterone supplementation of adult male rats suggested that none of the cloned mRNAs was directly androgen-regulated. Efferent duct ligation, however, showed that Re8 mRNA levels depended on testicular factors other than androgens. Conclusion: The novel rodent cDNAs can now be used to monitor epididymal gene expression more closely and to set up various regulatory and functional studies.展开更多
The function and usage of vMIPα encoded by K6 gene of herpesvirus 8 (HHV8) which has homology with human macrophage protein (MIP) have not been clearly known. In the present note the K6 gene of HHV8 was cloned and tr...The function and usage of vMIPα encoded by K6 gene of herpesvirus 8 (HHV8) which has homology with human macrophage protein (MIP) have not been clearly known. In the present note the K6 gene of HHV8 was cloned and transfected into NIH3T3 cells and E. coli cells. Conditional media from the 3T3-transfected cells and K6 product vMIPa from E. coli. Cells were used to perform the experiments of ligand-receptor binding and cellular adhesion with peripheral blood macrophages. The conditional media and the purified vMIPa from E. coli could compete to bind to CCR5 located on macrophages from peripheral blood with I125-hMIP-1α chemokine of human. Cellular adhesion showed that the conditional media from transfected cells and the purified vMIPa did not induce the adhesion of macrophages from peripheral blood to ICAM-1. In conclusion, vMIPα encoded by K6 gene of HHV8 can bind to CCR5 of peripheral blood macrophage cells and does not induce their adhesion. This suggests that vMIPa enclosed CCR5, also known as HIV展开更多
文摘目的·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敲低后转录上调基因附近的位点,其中包括BCAT1转录起始位点。结论·CBX8在人前列腺癌中高表达。CBX8具有抑制肿瘤细胞侵袭的功能。其机制可能是CBX8/PRC1复合体结合于BCAT1转录起始位点并抑制BCAT1转录。
文摘色素框同源蛋白8(chromobox protein homolog 8,CBX8)是PcG家族蛋白PRC1复合体的核心组成部分,在细胞增殖、衰老、维持干细胞自我更新和全能性及肿瘤发生中发挥重要作用.目前研究发现CBX8在多种恶性肿瘤中表达增高,并与肿瘤的进展及预后密切相关,已成为当前肿瘤领域的研究热点.本文就当前CBX8在肿瘤中的研究作一综述.
文摘<abstract>Aim: Identification of the rodent counterparts of human and canine epididymal cDNAs HE3, HE4 and Ce8/Ly6G5C by sequence homology and analysis of their expression patterns and regulation level in the rat. Methods: 'Electronic screening' of Expressed Sequence Tag (EST) and genomic databases, followed by RT-PCR and Northern blot analysis. Results: Rodent ESTs and genomic sequences homologous to HE3, HE4 and Ce8/Ly6G5C were identified in the public databases and the 'full-length' rat cDNAs cloned. To emphasise their homology to the human and canine genes, they were named Me3/Re3, Me4/Re4 and Re8 for mouse and rat counterparts, respectively. mRNA expression patterns were analysed in rats, including rat HEl and HE5/CD52 counterparts as controls. Re3 and Re8 mRNAs were only found in the rat epididymis, while Re4 showed a broader tissue distribution. Within the epididymis, Re3 and Re4 mRNAs were detected in all regions; Re8, on the other hand, was restricted to the caput. During postnatal development, Re3 and control mRNAs were found from the earliest stages investigated, while Re8 mRNA was observed only from day 24 postnatum, corresponding to the onset of spermatogenesis in the prepubertal testis. Castration and testosterone supplementation of adult male rats suggested that none of the cloned mRNAs was directly androgen-regulated. Efferent duct ligation, however, showed that Re8 mRNA levels depended on testicular factors other than androgens. Conclusion: The novel rodent cDNAs can now be used to monitor epididymal gene expression more closely and to set up various regulatory and functional studies.
基金This work was supported by the National "863" Program (Grant No. G0208070599) the Guangdong Provincial Natural Science Foundation (Grant No. 990470) and the National Natural Science Foundation of China (Grant No. 30070697).
文摘The function and usage of vMIPα encoded by K6 gene of herpesvirus 8 (HHV8) which has homology with human macrophage protein (MIP) have not been clearly known. In the present note the K6 gene of HHV8 was cloned and transfected into NIH3T3 cells and E. coli cells. Conditional media from the 3T3-transfected cells and K6 product vMIPa from E. coli. Cells were used to perform the experiments of ligand-receptor binding and cellular adhesion with peripheral blood macrophages. The conditional media and the purified vMIPa from E. coli could compete to bind to CCR5 located on macrophages from peripheral blood with I125-hMIP-1α chemokine of human. Cellular adhesion showed that the conditional media from transfected cells and the purified vMIPa did not induce the adhesion of macrophages from peripheral blood to ICAM-1. In conclusion, vMIPα encoded by K6 gene of HHV8 can bind to CCR5 of peripheral blood macrophage cells and does not induce their adhesion. This suggests that vMIPa enclosed CCR5, also known as HIV
