AIM: To isolate and identify differentially expressed proteins between cancer and normal tissues of gastric cancer by two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flig...AIM: To isolate and identify differentially expressed proteins between cancer and normal tissues of gastric cancer by two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). METHODS: Soluble fraction proteins of gastric cancer tissues and paired normal tissues were separated by 2-DE. The differentially expressed proteins were selected and identified by MALDI-TOF-MS and database search. RESULTS: 2-DE profiles with high resolution and reproducibility were obtained. Twenty-three protein spots were excised from sliver staining gel and digested in gel by trypsin, in which fifteen protein spots were identified successfully. Among the identified proteins, there were ten over-expressed and five under-expressed proteins in stomach cancer tissues compared with normal tissues. CONCLUSION: In this study, the well-resolved, reproducible 2-DE patterns of human gastric cancer tissue and paired normal tissue were established and optimized and certain differentially-expressed proteins were identified. The combined use of 2-DE and MS provides an effective approach to screen for potential tumor markers.展开更多
目的 分别建立肠型、弥漫型胃癌组织和血清双向凝胶电泳图谱,鉴定差异表达蛋白,从中发现有意义的胃癌肿瘤相关标志物.方法 采用双向凝胶电泳（two-dimensional polyacrylamide gel electrophoresis,2-DE）分离组织和血清总蛋白,经硝酸...目的 分别建立肠型、弥漫型胃癌组织和血清双向凝胶电泳图谱,鉴定差异表达蛋白,从中发现有意义的胃癌肿瘤相关标志物.方法 采用双向凝胶电泳（two-dimensional polyacrylamide gel electrophoresis,2-DE）分离组织和血清总蛋白,经硝酸银染色,分析选取有表达差异的蛋白质点,通过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）鉴定差异蛋白.结果 获得重复性和分辨率较好的胃癌组织和血清双向凝胶电泳图谱,经MALDI-TOF-MS鉴定得到20种蛋白,在7例（58.3%）肠型胃癌组织中均有硒结合蛋白1（SBP1）表达,KIAA基因家族蛋白在弥漫型胃癌组织和血清中均有表达.结论 建立了弥漫型肠型胃癌组织和血清双向凝胶电泳图谱,获得了差异蛋白质点,为寻找胃癌的特异蛋白质作为胃癌肿瘤标志物奠定了基础.展开更多
基金National High Technology Research and Development Program of China (863 Program) No. 2006AA 02A301 and No. 2007AA02Z179Science and Technology Commission of Shanghai Municipality, No. 07jc14041National Key Fundamental Research 973 Project, No. 2002CB13700
文摘AIM: To isolate and identify differentially expressed proteins between cancer and normal tissues of gastric cancer by two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). METHODS: Soluble fraction proteins of gastric cancer tissues and paired normal tissues were separated by 2-DE. The differentially expressed proteins were selected and identified by MALDI-TOF-MS and database search. RESULTS: 2-DE profiles with high resolution and reproducibility were obtained. Twenty-three protein spots were excised from sliver staining gel and digested in gel by trypsin, in which fifteen protein spots were identified successfully. Among the identified proteins, there were ten over-expressed and five under-expressed proteins in stomach cancer tissues compared with normal tissues. CONCLUSION: In this study, the well-resolved, reproducible 2-DE patterns of human gastric cancer tissue and paired normal tissue were established and optimized and certain differentially-expressed proteins were identified. The combined use of 2-DE and MS provides an effective approach to screen for potential tumor markers.
文摘目的 分别建立肠型、弥漫型胃癌组织和血清双向凝胶电泳图谱,鉴定差异表达蛋白,从中发现有意义的胃癌肿瘤相关标志物.方法 采用双向凝胶电泳（two-dimensional polyacrylamide gel electrophoresis,2-DE）分离组织和血清总蛋白,经硝酸银染色,分析选取有表达差异的蛋白质点,通过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）鉴定差异蛋白.结果 获得重复性和分辨率较好的胃癌组织和血清双向凝胶电泳图谱,经MALDI-TOF-MS鉴定得到20种蛋白,在7例（58.3%）肠型胃癌组织中均有硒结合蛋白1（SBP1）表达,KIAA基因家族蛋白在弥漫型胃癌组织和血清中均有表达.结论 建立了弥漫型肠型胃癌组织和血清双向凝胶电泳图谱,获得了差异蛋白质点,为寻找胃癌的特异蛋白质作为胃癌肿瘤标志物奠定了基础.
