CAJ | 학술논문

目的运用2-DE和MALDI-TOF-MS研究MM患者骨髓上清液中差异表达蛋白,以寻找对MM发病机制研究和诊断或鉴别诊断的特异性蛋白质标志物.方法收集14例MM患者、5例其他血液系统恶性肿瘤患者及5名健康对照者骨髓上清液标本.除去标本中白蛋白和免疫球蛋白G(IgG)后,用2-DE分离3组骨髓上清液标本.用ImageMaster 2D platinum 5.0图像分析软件分析比较3组2-DE凝胶图像,以差异倍数在3倍以上的蛋白点作为候选差异表达蛋白.然后,用MALDI-TOF-MS对重复性较好的候选差异表达蛋白做PMF和二级质谱鉴定.将得到的肽片段质量进行NCBInr 数据库检索,以鉴定出相应的差异表达蛋白.结果用图像分析软件对3组骨髓上清液2-DE凝胶进行分析比较,从MM组与健康对照组中筛选出47个差异蛋白点,从MM组与疾病对照组中筛选出58个差异蛋白点.从MM组选取41个差异点进行质谱分析并鉴定,发现MM组与其他2组相比,5种高表达蛋白为免疫球蛋白J链、κ轻链、λ轻链、前病毒遗传的Gag多聚蛋白和与半抗原结合的含成熟氧(末)端的催化抗体；3种低表达蛋白为血红蛋白、结合珠蛋白(Hp2)、锌-α2-糖蛋白.这些差异蛋白部分与MM的临床表现相关,部分与MM的发生发展及临床治疗相关.结论应用2-DE和MALDI-TOF-MS技术从MM患者骨髓上清液标本中筛出与MM疾病相关的8种差异表达蛋白,为进一步研究MM发生发展机制,完善MM诊断、鉴别诊断指标提供了参考依据.
목적 운용2-DE화MALDI-TOF-MS연구MM환자골수상청액중차이표체단백,이심조대MM발병궤제연구화진단혹감별진단적특이성단백질표지물.방법 수집14례MM환자、5례기타혈액계통악성종류환자급5명건강대조자골수상청액표본.제거표본중백단백화면역구단백G(IgG)후,용2-DE분리3조골수상청액표본.용ImageMaster 2D platinum 5.0도상분석연건분석비교3조2-DE응효도상,이차이배수재3배이상적단백점작위후선차이표체단백.연후,용MALDI-TOF-MS대중복성교호적후선차이표체단백주PMF화이급질보감정.장득도적태편단질량진행NCBInr 수거고검색,이감정출상응적차이표체단백.결과 용도상분석연건대3조골수상청액2-DE응효진행분석비교,종MM조여건강대조조중사선출47개차이단백점,종MM조여질병대조조중사선출58개차이단백점.종MM조선취41개차이점진행질보분석병감정,발현MM조여기타2조상비,5충고표체단백위면역구단백J련、κ경련、λ경련、전병독유전적Gag다취단백화여반항원결합적함성숙양(말)단적최화항체；3충저표체단백위혈홍단백、결합주단백(Hp2)、자-α2-당단백.저사차이단백부분여MM적림상표현상관,부분여MM적발생발전급림상치료상관.결론 응용2-DE화MALDI-TOF-MS기술종MM환자골수상청액표본중사출여MM질병상관적8충차이표체단백,위진일보연구MM발생발전궤제,완선MM진단、감별진단지표제공료삼고의거.
Objective To analyze the differentially expressed proteins in bone marrow supernatants of multiple myeloma patients by using 2-DE and MALDI-TOF-MS,and search for the special protein markers for studying the mechanism of the development and diagnosis or differential diagnosis of multiple myeloma.Methods The bone marrow supernatant samples of fourteen multiple myeloma patients,five other hematologic malignancies and five normal controls were collected.After removing albumin and IgG,the proteins in the supernatants were separated by 2-DE.Three groups images were analyzed and compared by Imagemnster 2D platinum 5.0 analysis software.Differentially expressed proteins were selected if the protein spots intensity showed more than 3 fold increase or decrease among different groups.The identities of the differentially expressed proteins with good repeatibility were determined by PMF based on by MALDI-TOF-MS or MALDI-TOF-MS/MS and NCBInr database search.Results 2-DE maps of bone marrow supernatants of the three groups could be analyzed and compared by image analysis of software.Forty-seven and fifty-eight differentially expressed protein spots were detected in multiple myeloma samples compared with normal controls and other hematologic malignancies samples,respectively.Forty-one reproducible spots were analyzed and identified by mass spectrum.Compared with other hematologic malignancies and normal controls,five up-expressed proteins and three down-expressed proteins were identified in multiple myeloma samples.They includes immunoglobulin J chain κ and λ light chain,provirus ancestral Gag polyprotein,mature oxy-cope catalytic antibody with hapten for up-expressed proteins,and hemoglobin,haptoglobin Hp2,zinc-alPha-2-glycoprotein for down-expressed proteins.These differentially expressed proteins reflect the features of muhiple myeloma,and relate to the development,progression and therapy of multiple myeloma.Conclusions Eight differentially expressed proteins in bone marrow supernatants of multiple myeloma are identified by using 2-DE and MALDI-TOF-MS.These differentially expressed proteins could be useful in studying the mechanism of the development and progression of multiple myeloma,and in developing diagnosis and differential diagnosis of multiple myeloma.