目的:应用激光显微切割(laser capture microdissection,LCM)结合蛋白组学技术筛查肝细胞癌(hepatocellular carcinoma,HCC)组织及其癌旁组织的差异表达蛋白,分析二甲基精氨酸二甲胺水解酶1(dimethylarginine dimethylaminohydrolase1,D...目的:应用激光显微切割(laser capture microdissection,LCM)结合蛋白组学技术筛查肝细胞癌(hepatocellular carcinoma,HCC)组织及其癌旁组织的差异表达蛋白,分析二甲基精氨酸二甲胺水解酶1(dimethylarginine dimethylaminohydrolase1,DDAH-1)在肝细胞癌表达的变化。方法:选取2003-2006年间东方肝胆外科医院原发性肝细胞癌患者的手术切除标本40例。利用LCM技术分离捕获肝癌组织及癌旁组织的肝实质细胞,应用二维凝胶电泳技术(two dimensional gel electrophoresis,2-DE)筛选全部标本共同差异表达频率>80%、差异强度>3倍的蛋白质点;应用电喷雾串联质谱(electrospray ionisation tandem mass spectrometry,ESI-MS/MS)及基质辅助激光解析电离飞行时间质谱(matrix-assisted laser desorption/ionization time of flight mass spectrometry,MALDI-TOF-MS)对差异蛋白点进行质谱鉴定分析;采用Western blotting及免疫组化对差异蛋白DDAH-1进行检测。结果:2-DE筛查获得在肝癌及癌旁组织差异表达的蛋白质点共20个,通过质谱鉴定获得12个蛋白质,其中5个在肝癌组织表达上调,7个下调;这12个蛋白质与细胞代谢、增殖、分化及信号调控相关,其中的DDAH-1是参与调控一氧化氮相关通路的重要酶。Western blotting检测显示,20例标本中16例DDAH-1表达明显升高;免疫组化检测证实,10例标本DDAH-1全部呈强阳性表达。结论:肝细胞癌组织细胞中筛查到12个差异表达蛋白质,其中的DDAH-1在肝癌组织中表达上调,其有可能在肝癌发生、发展过程中起重要作用。展开更多
为进一步了解冷驯化后的青稞苗对低温胁迫的响应机制,本研究采用藏青148作为材料,利用同位素相对标记与绝对定量技术(isobaric tags for relative and absolute quantitation, iTRAQ)标记联合液相色谱串联质谱(LC-MS/MS)技术并结合生物...为进一步了解冷驯化后的青稞苗对低温胁迫的响应机制,本研究采用藏青148作为材料,利用同位素相对标记与绝对定量技术(isobaric tags for relative and absolute quantitation, iTRAQ)标记联合液相色谱串联质谱(LC-MS/MS)技术并结合生物信息学和分子生物学技术对青稞低温诱导的蛋白组差异蛋白谱进行分析。结果表明,在iTRAQ测序分析的常温及冷驯化后低温处理的样品肽段图谱中,共鉴定出5111个蛋白质,肽链长度主要分布在6-24个氨基酸之间。通路分析显示,鉴定到的蛋白质被富集到126个通路。COG分析显示,R类蛋白数量最多,同时鉴定的蛋白涉及能量代谢、信号转导、防御机制等多个方面。对照常温和低温处理18h筛选到的差异蛋白有886个,存在499个上调,387个下调,且在代谢通路、乙醛酸代谢、三羧酸循环、糖酵解四条通路显著富集。展开更多
AIM: To preliminarily test proteomics in aqueous humor in patients with dry age-related macular degeneration(AMD) by using the proteomic technology.METHODS: Aqueous humor samples were collected from patients with or w...AIM: To preliminarily test proteomics in aqueous humor in patients with dry age-related macular degeneration(AMD) by using the proteomic technology.METHODS: Aqueous humor samples were collected from patients with or without dry AMD, who underwent cataract surgery. The aqueous samples were analyzed with isobaric tags for relative and absolute quantification(i TRAQ) combined with liquid chromatography tandem mass spectrometry(LC-MS/MS) technology. The differential expressed proteins were analyzed with gene ontology(GO) enrichment, Kyoto Encyclopedia of Genes and Genomes(KEGG) and protein-protein interaction(PPI) network analysis. The data were partly validated by ELISA and Western blot. False discovery rate(FDR) was used for statistical analysis. RESULTS: A total of 244 proteins were detected, in which 38 proteins were up-regulated and 51 were down-regulated significantly in patients with dry AMD compared with that in control groups(FDR value <1.0%). Several proteins, e.g., protein S100-A8(S10 A8), dystroglycan(DAG1), Ig alpha-1 chain C region(IGHA1), carbonic anhydrase 3(CAH3) and alpha-1-acid glycoprotein(A1 AG1) were increased more than 5 times of that in control group. The bioinformatics analysis showed that dry AMD is closely associated with inflammation or immune reaction, oxidative stress, blood coagulation and remodeling of extracellular matrix.CONCLUSION: i TRAQ-based proteomic analysis of aqueous humor demonstrate the differential expressions of proteins between dry AMD and control groups, providing the clues to understand the mechanisms and possible treatments of dry AMD.展开更多
文摘目的:应用激光显微切割(laser capture microdissection,LCM)结合蛋白组学技术筛查肝细胞癌(hepatocellular carcinoma,HCC)组织及其癌旁组织的差异表达蛋白,分析二甲基精氨酸二甲胺水解酶1(dimethylarginine dimethylaminohydrolase1,DDAH-1)在肝细胞癌表达的变化。方法:选取2003-2006年间东方肝胆外科医院原发性肝细胞癌患者的手术切除标本40例。利用LCM技术分离捕获肝癌组织及癌旁组织的肝实质细胞,应用二维凝胶电泳技术(two dimensional gel electrophoresis,2-DE)筛选全部标本共同差异表达频率>80%、差异强度>3倍的蛋白质点;应用电喷雾串联质谱(electrospray ionisation tandem mass spectrometry,ESI-MS/MS)及基质辅助激光解析电离飞行时间质谱(matrix-assisted laser desorption/ionization time of flight mass spectrometry,MALDI-TOF-MS)对差异蛋白点进行质谱鉴定分析;采用Western blotting及免疫组化对差异蛋白DDAH-1进行检测。结果:2-DE筛查获得在肝癌及癌旁组织差异表达的蛋白质点共20个,通过质谱鉴定获得12个蛋白质,其中5个在肝癌组织表达上调,7个下调;这12个蛋白质与细胞代谢、增殖、分化及信号调控相关,其中的DDAH-1是参与调控一氧化氮相关通路的重要酶。Western blotting检测显示,20例标本中16例DDAH-1表达明显升高;免疫组化检测证实,10例标本DDAH-1全部呈强阳性表达。结论:肝细胞癌组织细胞中筛查到12个差异表达蛋白质,其中的DDAH-1在肝癌组织中表达上调,其有可能在肝癌发生、发展过程中起重要作用。
文摘为进一步了解冷驯化后的青稞苗对低温胁迫的响应机制,本研究采用藏青148作为材料,利用同位素相对标记与绝对定量技术(isobaric tags for relative and absolute quantitation, iTRAQ)标记联合液相色谱串联质谱(LC-MS/MS)技术并结合生物信息学和分子生物学技术对青稞低温诱导的蛋白组差异蛋白谱进行分析。结果表明,在iTRAQ测序分析的常温及冷驯化后低温处理的样品肽段图谱中,共鉴定出5111个蛋白质,肽链长度主要分布在6-24个氨基酸之间。通路分析显示,鉴定到的蛋白质被富集到126个通路。COG分析显示,R类蛋白数量最多,同时鉴定的蛋白涉及能量代谢、信号转导、防御机制等多个方面。对照常温和低温处理18h筛选到的差异蛋白有886个,存在499个上调,387个下调,且在代谢通路、乙醛酸代谢、三羧酸循环、糖酵解四条通路显著富集。
基金Supported by National Natural Science Foundation of China(No.81570852)the Shanghai Municipal Health and Planning Commission Foundation(No.201540046)
文摘AIM: To preliminarily test proteomics in aqueous humor in patients with dry age-related macular degeneration(AMD) by using the proteomic technology.METHODS: Aqueous humor samples were collected from patients with or without dry AMD, who underwent cataract surgery. The aqueous samples were analyzed with isobaric tags for relative and absolute quantification(i TRAQ) combined with liquid chromatography tandem mass spectrometry(LC-MS/MS) technology. The differential expressed proteins were analyzed with gene ontology(GO) enrichment, Kyoto Encyclopedia of Genes and Genomes(KEGG) and protein-protein interaction(PPI) network analysis. The data were partly validated by ELISA and Western blot. False discovery rate(FDR) was used for statistical analysis. RESULTS: A total of 244 proteins were detected, in which 38 proteins were up-regulated and 51 were down-regulated significantly in patients with dry AMD compared with that in control groups(FDR value <1.0%). Several proteins, e.g., protein S100-A8(S10 A8), dystroglycan(DAG1), Ig alpha-1 chain C region(IGHA1), carbonic anhydrase 3(CAH3) and alpha-1-acid glycoprotein(A1 AG1) were increased more than 5 times of that in control group. The bioinformatics analysis showed that dry AMD is closely associated with inflammation or immune reaction, oxidative stress, blood coagulation and remodeling of extracellular matrix.CONCLUSION: i TRAQ-based proteomic analysis of aqueous humor demonstrate the differential expressions of proteins between dry AMD and control groups, providing the clues to understand the mechanisms and possible treatments of dry AMD.
