A novel HBV integration site involved in hepatocarcinogenesis was investigated. The HBV DNA integration sites were detected by Alu-PCR in hepatocellular carcinoma tissues, matched surrounding liver tissues in 30 patie...A novel HBV integration site involved in hepatocarcinogenesis was investigated. The HBV DNA integration sites were detected by Alu-PCR in hepatocellular carcinoma tissues, matched surrounding liver tissues in 30 patients with hepatitis B-related hepatocellular carcinoma (HCC) and 3 cases of normal liver tissues. The integration sites and flanking sequences in human genome were sequenced and blasted, and the expression of integrated HBV genes was determined by reverse transcriptase-polymerase chain reaction (RT-PCR). The influence of the up-regulated expression of integrated genes on hepatocarcinogenesis was analyzed. Nineteen integration sites of HBV DNA into HCC tissues were obtained by RT-PCR and sequencing. These genes encoding proteins were: LOC51030, LOC157777, minichromosome maintenance complex component 3 associated protein (MCM3AP), MCTP1, SH3 and multiple ankyrin repeat domains 2 isoform 2, CCDC40, similar to HCG2033532, mitochondrial ribosomal S5 pseudogene 4. One of them was integrated into the intron of MCM3AP. RT-PCR demonstrated that the expression levels of MCM3AP mRNA in HCC tissues, matched surrounding liver tissues and normal liver tissues were in a descendent order. The ratio of MCM3AP mRNA to the GAPDH mRNA in these three tissues was 1.07375, 0.21573, 0.06747 respectively, with the difference being statistically significant among them (P<0.05). Meanwhile, the expression levels of MCM3AP mRNA from HCC tissues in which HBV DNA integrated into MCM3AP were still significantly higher than those without HBV DNA integrated into MCM3AP. It was concluded that the HBV DNA integration sites into human genome were random, and MCM3AP was a new site. The up-regulated MCM3AP mRNA may affect flanking sequences which promote the hepatocarcinogenesis.展开更多
目的:分析微小染色体维持蛋白3(minichromosome maintenance protein 3,MCM3)在脑胶质瘤中的表达情况、临床意义和可能参与的生物学过程,并探究其与胶质瘤免疫的关系。方法:在线检索GEPIA和Oncomine数据库获得MCM3在胶质瘤组织中的表达...目的:分析微小染色体维持蛋白3(minichromosome maintenance protein 3,MCM3)在脑胶质瘤中的表达情况、临床意义和可能参与的生物学过程,并探究其与胶质瘤免疫的关系。方法:在线检索GEPIA和Oncomine数据库获得MCM3在胶质瘤组织中的表达情况,利用CGGA数据库在线分析MCM3表达和胶质瘤临床病理特征的关系。同时收集2019年1月到2020年3月在山西省人民医院神经外科接受手术治疗的24例胶质瘤患者的肿瘤标本和8例非肿瘤对照标本,采用免疫组化SP法检测MCM3的表达,对生物信息学分析结果进行验证。在TCGA和CGGA数据库中利用Kaplan-Meier生存曲线评价MCM3对胶质瘤预后的作用。通过Linkedomic数据库、STRING数据库和Cytoscape软件获得与MCM3表达显著相关的基因。使用DAVID数据库对MCM3及其显著相关基因进行GO和KEGG分析,探究基因功能。最后在TIMER数据库中探究MCM3表达和胶质瘤免疫浸润的关系。结果:综合生物信息学与临床数据分析显示,MCM3在胶质瘤组织中相对正常组织呈高表达(P=0.024),其表达量随着病理级别逐渐升高(P=0.001)。生存分析显示,MCM3高表达与胶质瘤不良预后有关(P<0.05)。GO和KEGG分析显示,MCM3及其显著相关基因主要富集于细胞周期、DNA复制和调节DNA损伤修复等方面。TIMER数据库分析结果显示,在胶质瘤队列中,MCM3与多种免疫浸润细胞具有相关性(P<0.05)。结论:MCM3在胶质瘤中高表达且与不良预后有关,其可能与胶质瘤细胞的细胞周期、DNA复制、调节DNA损伤修复和免疫微环境有关。MCM3能促进胶质瘤的进展,可作为胶质瘤患者预后判断指标和潜在的治疗靶点。展开更多
基金supported by grants from the National Natural Sciences Foundation of China (No. 30872237)the National Program on Key Basic Research Project (973 Program) (No. 2007CB512900)
文摘A novel HBV integration site involved in hepatocarcinogenesis was investigated. The HBV DNA integration sites were detected by Alu-PCR in hepatocellular carcinoma tissues, matched surrounding liver tissues in 30 patients with hepatitis B-related hepatocellular carcinoma (HCC) and 3 cases of normal liver tissues. The integration sites and flanking sequences in human genome were sequenced and blasted, and the expression of integrated HBV genes was determined by reverse transcriptase-polymerase chain reaction (RT-PCR). The influence of the up-regulated expression of integrated genes on hepatocarcinogenesis was analyzed. Nineteen integration sites of HBV DNA into HCC tissues were obtained by RT-PCR and sequencing. These genes encoding proteins were: LOC51030, LOC157777, minichromosome maintenance complex component 3 associated protein (MCM3AP), MCTP1, SH3 and multiple ankyrin repeat domains 2 isoform 2, CCDC40, similar to HCG2033532, mitochondrial ribosomal S5 pseudogene 4. One of them was integrated into the intron of MCM3AP. RT-PCR demonstrated that the expression levels of MCM3AP mRNA in HCC tissues, matched surrounding liver tissues and normal liver tissues were in a descendent order. The ratio of MCM3AP mRNA to the GAPDH mRNA in these three tissues was 1.07375, 0.21573, 0.06747 respectively, with the difference being statistically significant among them (P<0.05). Meanwhile, the expression levels of MCM3AP mRNA from HCC tissues in which HBV DNA integrated into MCM3AP were still significantly higher than those without HBV DNA integrated into MCM3AP. It was concluded that the HBV DNA integration sites into human genome were random, and MCM3AP was a new site. The up-regulated MCM3AP mRNA may affect flanking sequences which promote the hepatocarcinogenesis.
文摘目的:分析微小染色体维持蛋白3(minichromosome maintenance protein 3,MCM3)在脑胶质瘤中的表达情况、临床意义和可能参与的生物学过程,并探究其与胶质瘤免疫的关系。方法:在线检索GEPIA和Oncomine数据库获得MCM3在胶质瘤组织中的表达情况,利用CGGA数据库在线分析MCM3表达和胶质瘤临床病理特征的关系。同时收集2019年1月到2020年3月在山西省人民医院神经外科接受手术治疗的24例胶质瘤患者的肿瘤标本和8例非肿瘤对照标本,采用免疫组化SP法检测MCM3的表达,对生物信息学分析结果进行验证。在TCGA和CGGA数据库中利用Kaplan-Meier生存曲线评价MCM3对胶质瘤预后的作用。通过Linkedomic数据库、STRING数据库和Cytoscape软件获得与MCM3表达显著相关的基因。使用DAVID数据库对MCM3及其显著相关基因进行GO和KEGG分析,探究基因功能。最后在TIMER数据库中探究MCM3表达和胶质瘤免疫浸润的关系。结果:综合生物信息学与临床数据分析显示,MCM3在胶质瘤组织中相对正常组织呈高表达(P=0.024),其表达量随着病理级别逐渐升高(P=0.001)。生存分析显示,MCM3高表达与胶质瘤不良预后有关(P<0.05)。GO和KEGG分析显示,MCM3及其显著相关基因主要富集于细胞周期、DNA复制和调节DNA损伤修复等方面。TIMER数据库分析结果显示,在胶质瘤队列中,MCM3与多种免疫浸润细胞具有相关性(P<0.05)。结论:MCM3在胶质瘤中高表达且与不良预后有关,其可能与胶质瘤细胞的细胞周期、DNA复制、调节DNA损伤修复和免疫微环境有关。MCM3能促进胶质瘤的进展,可作为胶质瘤患者预后判断指标和潜在的治疗靶点。
