Protein ubiquitination is essential for diverse cellular functions including spermatogenesis.The tripartite motif(TRIM)family proteins,most of which have E3 ubiquitin ligase activity,are highly conserved in mammals.Th...Protein ubiquitination is essential for diverse cellular functions including spermatogenesis.The tripartite motif(TRIM)family proteins,most of which have E3 ubiquitin ligase activity,are highly conserved in mammals.They are involved in important cellular processes such as embryonic development,immunity,and fertility.Our previous studies indicated that Trim69,a testis-specific expressed TRIM family gene,potentially participates in the spermatogenesis by mediating testicular cells apoptosis.In this study,we investigated the biological functions of Trim69 in male mice by established Trim69 knockout mice with CRISPR/Cas9 genomic editing technology.Here,we reported that the male Trim69 knockout mice had normal fertility.The adult knockout mice have shown that the appearance of testes,testis/body weight ratios,testicular histomorphology,and the number and quality of sperm were consistent with wild-type mice.These results indicated that the E3 ubiquitin ligase protein Trim69 was not essential for male mouse fertility,and it might be compensated by other TRIM family members such as Trim58 in Trim69-deficiency testis.This study would help to elucidate the functions of tripartite motif protein family and the regulation of spermatogenesis.展开更多
目的研究根皮苷对食管癌细胞系表达谱的影响,探索根皮苷作用于食管癌细胞系的信号通路及其潜在功能。方法使用根皮苷处理人食管癌KYSE450细胞,提取总RNA并建立测序文库,进行转录组测序;使用DESeq2程序鉴定差异表达基因,对差异表达基因...目的研究根皮苷对食管癌细胞系表达谱的影响,探索根皮苷作用于食管癌细胞系的信号通路及其潜在功能。方法使用根皮苷处理人食管癌KYSE450细胞,提取总RNA并建立测序文库,进行转录组测序;使用DESeq2程序鉴定差异表达基因,对差异表达基因进行基因本体(gene ontology,GO)功能及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;使用MCODE进行差异基因核心模块筛选,使用GEPIA数据库分析核心模块中的基因对食管癌生存的影响;使用TIMER在线分析根皮苷对食管癌组织免疫细胞浸润的影响。结果转录组测序结果显示,根皮苷处理后的食管癌细胞具有4602个差异表达基因,其中2407个为上调基因,2195个为下调基因。通路富集分析显示,根皮苷对蛋白质加工、胰岛素抵抗、基因复制和细胞周期等信号通路产生影响。在差异表达基因核心模块中,E3泛素蛋白连接酶2(E3 ubiquitin ligase mind bomb 2,MIB2)高表达可降低食管癌患者的总体生存率;环指蛋白19B(ring finger protein19B,RNF19B)、三重基序蛋白69(tripartite motif-containing protein 69,TRIM69)、泛素连接酶(ubiquitin conjugating enzyme,UBC)和克隆E3泛素连接酶(homologous to the E6-associated protein carboxyl terminus domain containing 2,HECTD2)的表达水平可影响食管癌癌组织的纯度以及免疫细胞浸润的类型。结论根皮苷可影响食管癌细胞的转录谱,其差异表达基因主要集中在细胞生长相关的信号通路。展开更多
基金supported by the National Key Research and Development Program of China(No.2018YFC1003503-2,No.2017YFA0103803,and No.2016YFA0500903)the National Natural Science Foundation of China(No.31890784)。
文摘Protein ubiquitination is essential for diverse cellular functions including spermatogenesis.The tripartite motif(TRIM)family proteins,most of which have E3 ubiquitin ligase activity,are highly conserved in mammals.They are involved in important cellular processes such as embryonic development,immunity,and fertility.Our previous studies indicated that Trim69,a testis-specific expressed TRIM family gene,potentially participates in the spermatogenesis by mediating testicular cells apoptosis.In this study,we investigated the biological functions of Trim69 in male mice by established Trim69 knockout mice with CRISPR/Cas9 genomic editing technology.Here,we reported that the male Trim69 knockout mice had normal fertility.The adult knockout mice have shown that the appearance of testes,testis/body weight ratios,testicular histomorphology,and the number and quality of sperm were consistent with wild-type mice.These results indicated that the E3 ubiquitin ligase protein Trim69 was not essential for male mouse fertility,and it might be compensated by other TRIM family members such as Trim58 in Trim69-deficiency testis.This study would help to elucidate the functions of tripartite motif protein family and the regulation of spermatogenesis.
文摘目的研究根皮苷对食管癌细胞系表达谱的影响,探索根皮苷作用于食管癌细胞系的信号通路及其潜在功能。方法使用根皮苷处理人食管癌KYSE450细胞,提取总RNA并建立测序文库,进行转录组测序;使用DESeq2程序鉴定差异表达基因,对差异表达基因进行基因本体(gene ontology,GO)功能及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;使用MCODE进行差异基因核心模块筛选,使用GEPIA数据库分析核心模块中的基因对食管癌生存的影响;使用TIMER在线分析根皮苷对食管癌组织免疫细胞浸润的影响。结果转录组测序结果显示,根皮苷处理后的食管癌细胞具有4602个差异表达基因,其中2407个为上调基因,2195个为下调基因。通路富集分析显示,根皮苷对蛋白质加工、胰岛素抵抗、基因复制和细胞周期等信号通路产生影响。在差异表达基因核心模块中,E3泛素蛋白连接酶2(E3 ubiquitin ligase mind bomb 2,MIB2)高表达可降低食管癌患者的总体生存率;环指蛋白19B(ring finger protein19B,RNF19B)、三重基序蛋白69(tripartite motif-containing protein 69,TRIM69)、泛素连接酶(ubiquitin conjugating enzyme,UBC)和克隆E3泛素连接酶(homologous to the E6-associated protein carboxyl terminus domain containing 2,HECTD2)的表达水平可影响食管癌癌组织的纯度以及免疫细胞浸润的类型。结论根皮苷可影响食管癌细胞的转录谱,其差异表达基因主要集中在细胞生长相关的信号通路。
