非小细胞肺癌分泌蛋白的蛋白质组学研究(英文)  

Proteomic Analysis of Secreted Proteins of Non-small Cell Lung Cancer

作　　者：黄凌瑾[1] 陈胜喜[1] 罗万俊[1] 蒋海河[1] 张鹏飞[2] 易红[2] 

机构地区：[1]中南大学湘雅医院心胸外科,湖南长沙410008 [2]中南大学湘雅医院医学研究中心,湖南长沙410008

出　　处：《癌症》2006年第11期1361-1367,共7页Chinese Journal of Cancer

摘　　要：背景与目的:癌细胞分泌蛋白是潜在的血清标志物,而且对肿瘤的早期筛选也具有重要意义。因此,系统、全面地研究肿瘤细胞分泌蛋白对于肿瘤血清标志物的筛选具有重要意义。本研究旨在鉴定肺癌细胞分泌蛋白,为肺癌血清标志物筛选提供实验依据。方法:用含胎牛血清的RPMI-1640培养液培养非小细胞肺癌细胞系A549细胞,收集A549细胞培养液中的蛋白及胎牛血清培养液中的蛋白(对照)进行蛋白双向电泳,应用基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)及生物信息学鉴定A549凝胶上特有的蛋白质点。筛选出人属的蛋白质点后,应用RT-PCR方法检测其在肺癌组织及配对远处肺组织中的表达差异;并检测培养液及血清中锰-超氧化物歧化酶(manganesesuperoxidedismutase,Mn-SOD)水平。结果:鉴定出人属的蛋白质点14个,包括肌动蛋白、!烯醇酶(alphaenolase,ENO1)、Mn-SOD、谷胱苷肽S转移酶P(glutathioneS-transferaseP,GSTP1-1)、二氢二醇脱氢酶(dihydrodioldehydrogenase,DDH)、磷酸甘油酸变位酶1(phosphoglyceratemutase1,PGAM1)、葡萄糖依赖性胰岛素释放肽受体(glucose-dependentinsulinotropicproteinreceptor,GIPR)、肽基脯氨基顺反异构酶(peptidyl-prolylcis-transisomeraseA,PPIA)、磷脂酰乙醇胺结合蛋白(phosphatidylethanolaminebindingprotein,PEBP)、蛋白基因产物9.5(ubiquitincarboxyl-terminalhydrolaseisozymeL1,PGP9.5)、peroxiredoxin1(PDX1)、galectin-1及专利蛋白WO0222660等。RT-PCR检测示ENO1、DDH、PEBP、PGP9.5、PDX1、PGAM1、PPIA在肺癌组织中高表达。酶活性检测发现A549培养液中存在Mn-SOD,且发现非小细胞肺癌患者血清中有高水平的Mn-SOD。结论:本研究首次对非小细胞肺癌细胞的分泌蛋白进行了鉴定;发现多种高表达的非小细胞肺癌分泌蛋白基因,其中首次发现ENO1及PEBP基因在非小细胞肺癌中高表达;Mn-SOD是非小细胞肺癌的分泌型血清标志物。该研究为非小细胞肺癌BACKGROUND ＆ OBJECTIVE： Secreted proteins from cancer ceils may be potential serologic biomarkers of cancer. It＇s important to globally identify secreted proteins of cancer cells. This study was to identify secreted proteins of lung cancer cells. METHODS： Proteins in the conditioned medium of non-small cell lung cancer （NSCLC） cell line A549 was collected and the proteome analysis was subsequently performed. Specific protein spots in A549 cells were identified by peptide mass fingerprints using mass spectrometry and through searching database. The expression of identified secreted proteins was detected by reverse transcription-polymerase chain reaction （RT-PCR） in 15 specimens of NSCLC tissue and paired distant lung tissue. Manganese superoxide dismutase （Mn-SOD） activity in serum and conditioned medium was detected by spectrophotometry. RESULTS： Fourteen secreted proteins were identified, which included peptidyl-prolyl cis-trans isomerase A （PPIA）, Mn- SOD, peroxiredoxin 1 （PDX1）, phosphatidylethanolamine binding protein （PEBP）, glutathione S-transferase P （GSTPI-1）, glucose-dependent insulinotropic protein receptor （GIPR）, ubiquitin carboxyl-terminal hydrolase isozyme L1 （PGP9.5）, alpha enolase （ENO1）, dihydrodiol dehyclrogenase （DDH）, phosphoglycerate mutase 1 （PGAM1）,galectin-1 （GALl）. PPIA, DDH, PGAM1, PDX1, PGP9.5, ENO1, and PEBP were overexpressed in cancer tissues. Higher level of Mn-SOD activity was detected in conditioned medium than in control. Serum Mn-SOD activity was significantly higher in NSCLC patients than in healthy controls （P〈0.01）. CONCLUSIONS. Multiple secreted proteins of A549 cells were identified in this study and the overexpression of ENO1 and PEBP in NSCLC was revealed for the first time. Mn-SOD is secreted serologic marker of NSCLC. The results presented here would provide clues to identify new serologic biomarkers of NSCLC.

关 键 词：肺肿瘤 非小细胞性 分泌蛋白 血清标志物 蛋白质组学 

分 类 号：R734.2[医药卫生—肿瘤]