利用血清蛋白质组差异分析多发性骨髓瘤的耐药机制

目的应用弱阳离子交换纳米磁珠(WCX-MB)分离血清蛋白并联合基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)技术,对比分析临床上难治复发与治疗敏感的多发性骨髓瘤(MM)患者血清蛋白质组的差异,以期为阐明多发性骨髓瘤的耐药机制提供线索。方法选择临床确诊的19例MM患者,按实际疗效分为难治复发组(11例)和治疗敏感组(8例),以WCX-MB捕获各自的血清蛋白质组分,MALDI-TOF-MS检测蛋白质谱,再利用ClinprotoolsTM 2.2软件进行数据分析,筛选出与耐药相关的差异蛋白质分子。结果在分子量0.7～10ku的范围内,共得到59个有统计学意义的差异多肽峰(P<0.05);与治疗敏感组相比,难治复发组MM患者的血清多肽峰有34个表达上调,有25个表达下调;将区分难治复发组与治疗敏感组MM患者能力最强的10个质谱峰建成了一个诊断GA模型,其灵敏度及特异性均为100%。结论利用CLINPROT系统筛选血清差异表达蛋白能够为研究多发性骨髓瘤的耐药机制和临床预后判定提供有价值的线索。

Mechanism of drug resistance in patients with multiple myeloma analyzed with serum proteome differencing

Objective To provide clues for studying the mechanism of drug resistance in patients with multiple myeloma(MM),the serum protein components of refractory and therapy susceptible patients were captured by weak cation exchange nanometer magnetic beads(WCX-MB) and the protein mass-spectra of all samples were detected by Autoflex II matrix-assisted laser desorption/ionization time of flight mass spectrometer(MALDI-TOF-MS) so as to compare their differences.Methods Altogether 19 patients with MM were enrolled in this study,including 11 cases of refractory or relapsed MM and 8 cases of therapy susceptible according to their clinical outcomes.The serum proteins were captured by WCX-MB and protein mass-spectra data detected by MALDI-TOF-MS were analyzed with ClinprotoolsTM 2.2 software to find the drug-resistance related proteins.Results Totally 59 significantly differential protein peaks(P<0.05) could be detected within the range of 0.7-10 ku in protein spectra of serum samples.Compared with therapy susceptible group,34 protein peaks were up-regulated and 25 protein peaks were down-regulated in refractory or relapsed MM patients.Diagnostic model was established on the basis of Genetic Algorithm(GA),and the 10 mass peaks had the strongest power to automatically distinguish susceptible group from refractory or relapsed group.Sensitivity and specificity were expected to be both 100% in this established model group.Conclusion Screening the serum differentially expressing proteins with CLINPROT system could provide valuable clues for analyzing the mechanism of drug resistance and prognosis determination in MM patients.