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1、SELDI技术分析体外培养不同肝细胞株差异蛋白的表达(1) 作者:丁守怡,钱冬萌,牟文凤,闫志勇,宋旭霞,王斌 【关键词】表面增强激光解吸/离子化/飞行时间质谱;蛋白质陈列分析;肝肿瘤;肿瘤细胞,培养的;差异蛋白 AnalysisofdifferentproteinexpressionsindifferentlivercellstrainsinvitrousingSELDI 【Abstract】AIM:Toexaminethedifferentialexpressionsofproteinsinhepatomac
2、ellline,hepatomacelllinetransfectedbyHBV()comparedwithnormallivercelllinebyusingsurfaceenhancedlaserdesorption/ionizationtimeofflightmassspectrometry,soastoestablishafoundationforfurtherstudyingonthemechanismofhepatomaattheproteinlevel.METHODS:The3typesofcellsabo
3、vementionedwereculturedasgeneralandcollectedwhentheywereingoodconditions.Aftercelldisruption,SELDITOFMSwasemployedtodetectthedifferentialexpressionsofproteomesofHepG2,HepGandLO2.RESULTS:NinetyoneproteinswerecapturedbyWCX2array.Comparedwithnormallivercelllines,int
4、hesegmentsfromMr5000to15000,proteinsintheHepG2andshowedapparentchanges,inwhich2proteinsexpressionswereincreasedincarcinomacells,theother5decreased.NineproteinsinHepG2wereincreasedand10proteinsinHepGwereincreased.CONCLUSION:TheSELDIProteinChiptechnologycanbeadesir
5、edmethodindetectingthebiologicalmarkersintheearlierperiodofhepatoma.Thismethodisofconvenience,highsensitivity,andgoodreproducibility.Thebiologicaltagsoftissuespecificproteinswefoundhaveapotentialapplyingvalueintheearlydiagnosisofhepatoma.Thesetagsarealsomeaningfu
6、linscreeningandidentifyingsignalproteinsfromserumandtissuespecimenindifferenttypesofhepatoma.Certainfoundationhasbeenestablishedinstudyingtheetiopathogenesisofhepatomaatthelevelofproteins,aswellasthesearchingofnewtherapeutictargetsites. 【Keywords】SELDITOFMS;prot
7、einarrayanalysis;liverneoplasms;tumorcells,cultured;distinctprotein 【摘要】目的:运用表面增强激光解吸/离子化/飞行时间质谱技术,检测体外培养的肝癌细胞株(HepG2),转染乙肝病毒的肝癌细胞株()与正常肝细胞株(LO2)蛋白质的差异表达,筛选肝细胞癌的标志蛋白,为进一步研究肝癌发病的蛋白质组学机制奠定基础.方法:常规培养上述3种细胞,细胞状态良好时收集细胞,裂解细胞后采用SELDITOFMS技术用WCX2芯片检测HepG2,,LO2细胞内蛋白质组学的
8、差异表达.结果:WCX2芯片共捕获91个蛋白,在Mr5000~15000区段,与正常肝细胞株比较,有7个蛋白质在两种肝癌细胞中出现明显变化,其中2个蛋白在肝癌细胞中表达量增高,5个蛋白在肝癌细胞中表达量降低;另外两种肝癌细胞株也有其各自的标志蛋白,9个蛋白分子在HepG2表达量增高,10个蛋白分子在表达量增高.结论: