目的:探究正常肺组织、肺腺癌癌旁和肺腺癌中所有基因的表达水平趋势,寻找特征性基因并进行实验验证。方法:下载加州大学圣克鲁斯分校(University of California,Santa Cruz,UCSC)数据中心中癌症基因组图谱(the cancer genome atlas,TC...目的:探究正常肺组织、肺腺癌癌旁和肺腺癌中所有基因的表达水平趋势,寻找特征性基因并进行实验验证。方法:下载加州大学圣克鲁斯分校(University of California,Santa Cruz,UCSC)数据中心中癌症基因组图谱(the cancer genome atlas,TCGA)和基因型-组织表达项目(genotype-tissue expression,GTEX)的整合数据,采用R语言分析所有基因的表达趋势,寻找特征性基因,提取癌旁和肿瘤组织共同高表达的基因集,进行GO和KEGG分析富集情况,鉴定最显著的通路中在肺组织和癌旁及肺腺癌中变化最大的基因。通过细胞生物学实验验证候选基因在肿瘤中的作用。结果:整合数据显示存在一群正常组织中低表达但在肺腺癌癌旁和肺腺癌中高表达的基因(DEGS2、PIGY、FUCA、GM2A),GO分析显示该基因群富集最显著的通路是膜脂质代谢过程,对该通路基因和符合趋势的基因取交集,发现DEGS2在癌旁和癌组织中升高最明显,升高倍数达3.27倍。实验证实,敲除DEGS224 h后细胞迁移能力下降,72 h后细胞活性下降55.0%。结论:DEGS2在癌旁和肺腺癌中呈高表达,其表达能促进肺腺癌的增殖和迁移,从而进一步驱动从癌旁组织到癌的发展。展开更多
The Synechocystis sp. PCC 6803 genome harbours a Deg gene family consisting of three members, htrA （degP, slr1204）, hhoA （degQ, sll1679） and hhoB （degS, sll1427）. This work provided biochemical characterization ...The Synechocystis sp. PCC 6803 genome harbours a Deg gene family consisting of three members, htrA （degP, slr1204）, hhoA （degQ, sll1679） and hhoB （degS, sll1427）. This work provided biochemical characterization of HhoA, HtrA and HhoB from Synechocystis sp. PCC 6803. Firstly mature HhoA, HhoB and HtrA from Synechocystis sp. PCC 6803 were cloned and expressed as soluble recombinant his-tagged fusion protein in Escherichia coli. Then the proteolytic activity of HhoA, HhoB and HtrA was tested using casein, bovine serum albumin, five recombinant chromoproteins and cyanobacterial phycocyanin as substrates in vitro. The experimental results showed that HhoA and HtrA had proteolytic activity on casein, five recombinant chromoproteins and cyanobacterial phycocyanin. No proteolytic activity of HhoB was found using all substrates in vitro, indicating functional difference among Deg proteases from Synechocystis sp. PCC 6803. Therefore, the results indicated the biochemical properties of HhoA and HtrA on hydrolysis of proteins and phycobiliproteins in vitro, which implicated that they were proteases possibly involved in phycobiliprotein degradation in vivo.展开更多
文摘目的:探究正常肺组织、肺腺癌癌旁和肺腺癌中所有基因的表达水平趋势,寻找特征性基因并进行实验验证。方法:下载加州大学圣克鲁斯分校(University of California,Santa Cruz,UCSC)数据中心中癌症基因组图谱(the cancer genome atlas,TCGA)和基因型-组织表达项目(genotype-tissue expression,GTEX)的整合数据,采用R语言分析所有基因的表达趋势,寻找特征性基因,提取癌旁和肿瘤组织共同高表达的基因集,进行GO和KEGG分析富集情况,鉴定最显著的通路中在肺组织和癌旁及肺腺癌中变化最大的基因。通过细胞生物学实验验证候选基因在肿瘤中的作用。结果:整合数据显示存在一群正常组织中低表达但在肺腺癌癌旁和肺腺癌中高表达的基因(DEGS2、PIGY、FUCA、GM2A),GO分析显示该基因群富集最显著的通路是膜脂质代谢过程,对该通路基因和符合趋势的基因取交集,发现DEGS2在癌旁和癌组织中升高最明显,升高倍数达3.27倍。实验证实,敲除DEGS224 h后细胞迁移能力下降,72 h后细胞活性下降55.0%。结论:DEGS2在癌旁和肺腺癌中呈高表达,其表达能促进肺腺癌的增殖和迁移,从而进一步驱动从癌旁组织到癌的发展。
基金Funded by the National Natural Science Foundation of China (Nos.30870541,30870519)
文摘The Synechocystis sp. PCC 6803 genome harbours a Deg gene family consisting of three members, htrA （degP, slr1204）, hhoA （degQ, sll1679） and hhoB （degS, sll1427）. This work provided biochemical characterization of HhoA, HtrA and HhoB from Synechocystis sp. PCC 6803. Firstly mature HhoA, HhoB and HtrA from Synechocystis sp. PCC 6803 were cloned and expressed as soluble recombinant his-tagged fusion protein in Escherichia coli. Then the proteolytic activity of HhoA, HhoB and HtrA was tested using casein, bovine serum albumin, five recombinant chromoproteins and cyanobacterial phycocyanin as substrates in vitro. The experimental results showed that HhoA and HtrA had proteolytic activity on casein, five recombinant chromoproteins and cyanobacterial phycocyanin. No proteolytic activity of HhoB was found using all substrates in vitro, indicating functional difference among Deg proteases from Synechocystis sp. PCC 6803. Therefore, the results indicated the biochemical properties of HhoA and HtrA on hydrolysis of proteins and phycobiliproteins in vitro, which implicated that they were proteases possibly involved in phycobiliprotein degradation in vivo.
