摘要
目的探讨胰腺癌的发病机制,寻找与临床诊断及治疗相关的生物学标记或靶标。方法采用二维聚丙烯酰胺凝胶电泳(2-DE)技术对5例人胰腺癌组织及其癌旁组织的总蛋白进行分离,PDQuest 7.0软件分析获得差异蛋白质点,基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)对蛋白质点进行鉴定,采用免疫组化染色进一步验证差异表达蛋白。结果共获得10个差异表达蛋白质点。质谱鉴定结果显示,尼克酰胺N-甲基转移酶(NNMT)和铁蛋白在胰腺癌组织中表达明显高于其在癌旁组织中表达(>4倍),而谷胱甘肽S-转移酶、DNA拓扑异构酶、T细胞受体β-链、IKK抑制因子ε1、转胶蛋白、二甲基精氨酸二甲基氨基水解酶1(DDAH1)、半乳糖激酶1、肌球蛋白等在胰腺癌癌旁组织中表达明显高于其在癌组织中表达(>4倍)。免疫组化结果显示,铁蛋白在胰腺癌组织中表达高于癌旁组织,DNA拓扑异构酶、谷胱甘肽S-转移酶、肌球蛋白在癌旁组织中表达高于癌组织。结论以2-DE和MALDI-TOF-MS为基础的蛋白质组学技术是研究肿瘤的一种重要手段,实验得到的差异蛋白可能是潜在诊断胰腺癌的分子标志或控制肿瘤生长的治疗靶点。
Objective To explore pathogenesis of pancreatic cancer and screen specific biological markers of clinical diagnosis and treatment associated with pancreatic cancer. Methods Total proteins of pancreatic cancer and adjacent normal tissues were obtained from five pancreatic cancer patients.Two-dimensional gelelectrophoresis（2-DE）,PDQuest 7.0 and MALDI-TOF-MS technique were used to get differential expression proteins of pancreatic cancer from adjacent normal tissues.Immunohistochemistry method was further performed to verify differential proteins. Results Ten differential proteins were obtained,including the two up-regulated proteins of nicotinamide N-methyltransferase（NNMT） and ferritin（〉4 folds） and the eight down-regulated proteins of glutathione S-transferases,DNA topoisomerase,T-cell receptor beta chain,suppressor of IKK epsilon isoform 1,transgelin,dimethylarginine dimethylamin-ohydrolase 1（DDAH1）,galactokinase 1,myosin（〉4 folds） in pancreatic tumor tissues.By immunohistochemistry,ferritin was up-regulated in tumor tissues,and DNA topoisomerase,glutathione S-transferases and myosin were up-regulated in normal tissues. Conclusion The analysis of proteomics with 2-DE and MALDI-TOF-MS method on human tissue is a useful method for discovering valuable cancer maker candidates.These differential expressed proteins may serve as biomarkers for early detection and therapeutic targets in pancreatic cancer.
出处
《临床肿瘤学杂志》
CAS
2011年第11期961-965,共5页
Chinese Clinical Oncology
基金
国家自然科学基金资助项目(30770612)
