【作者】 张巧花； 侯淑玲； 王军； 贺建霞； 郭素堂；

【机构】 山西省肿瘤医院血液内科； 山西省肿瘤研究所；

【摘要】 目的:应用表面增强激光解吸／离子化-飞行时间- 质谱(surface enhanced laser desorption／ionization—time of flight—mass spectrometry,SELDI-TOF-MS)蛋白质芯片技术检测B细胞非霍奇金淋巴瘤(B—cell non-Hodgkin lymphoma,B-cell NHL)及其弥漫大B细胞淋巴瘤 (diffuse large B—cell lymphoma,DLBCL)和滤泡性淋巴瘤(follicular lymphoma,FL)病人血清中的差异表达蛋白, 并在化疗后对其进一步检测以探讨其临床意义。方法:应用 SELDI-TOF—MS技术通过弱阳离子交换(WCX-2)蛋白芯片检测54例初治B-cell NHL患者和27例正常人血清中的差异表达蛋白;同时筛选其病理类型——23例DLBCL患者和12例FL患者之间的差异表达蛋白,以及影响23例DLBCL 患者预后的差异表达蛋白。结果: (1)采用PBS-II型蛋白质芯片阅读机,自动收集蛋白质峰,于54例B-cell NHL患者和27例正常人血清中在相对分子量3000—20000Da范围内检测出相对高丰度的6个差异表达蛋白,其相对分子量为 7974Da、15938Da、3398Da、8564Da、8692Da和3193Da。经Biomarker Wizard软件分析在54例B-cell NHL患者和27例正常人血清中有差异,有显著的统计学意义。 (2) WCX-2蛋白质芯片检测的结果表明,与正常对照组血清蛋白质谱相比,B—cell NHL病人血清中有2个高表达的差异表达蛋白,它们的相对分子量分别为7974Da和15938Da,经 Biomarker Wizard软件分析在54例B-cell NHL患者和27 例正常人血清中有差异,有显著的统计学意义。(3)WCX-2 蛋白质芯片检测的结果表明,与正常对照组血清蛋白质谱相比,B-cell NHL病人血清中有4个低表达的差异表达蛋白, 它们的相对分子量分别为:3398Da、8564Da、8692Da和 3193Da。经Biomarker Wizard软件分析在54例B-cell NHL患者和27例正常人血清中有差异,有显著的统计学意义。(4)WCX-2蛋白质芯片检测的结果表明,DLBCL中低表达且FL中高表达的差异表达蛋白的相对分子量分别为 3379Da、3976Da和4302Da。经Biomarker Wizard软件分析有显著的统计学意义。(5)WCX-2蛋白质芯片检测的结果表明,其中11例DLBCL长期缓解(完全缓解期>6个月)、 12例DLBCL预后不良(达不到完全缓解和完全缓解期 6 个月),其二者间差异表达蛋白为4795Da和4998Da,其高表达时DLBCL患者长期缓解,经Biomarker Wizard软件分析有显著的统计学意义。(6)27例初治B—cell NHL患者经二周期化疗(BACOP方案或COEP方案)均达到了部分缓解或完全缓解。此时,用WCX-2芯片检测化疗后的血清中差异表达蛋白,同样用PBS-II型蛋白质芯片阅读机,自动采集蛋白质峰,结果表明B-cell NHL患者血清中高表达的差异表达蛋白化疗病情好转后出现低表达,B—cell NHL患者血清中低表达的差异表达蛋白化疗病情好转后出现高表达, 经配对资料的t-检验,相对分子量为15938Da、7974Da、 3398Da、8564Da和8692Da的蛋白质具有统计学意义;相对分子量为3193Da的蛋白质没有统计学意义。结论:B—cell NHL的标志蛋白:相对分子量为7974Da、15938Da、3398Da、 8564Da和8692Da,其中7974Da和15938Da高表达,3398Da、 8564Da和8692Da低表达。DLBCL与FL之间有三个差异表达蛋白,其相对分子量为3379Da、3976Da和4302Da,它们在FL中高表达,在DLBCL中低表达。有两个差异表达蛋白影响DLBCL的预后,相对分子量为4795Da和4998Da,其高表达时化疗后可以长期缓解。血清蛋白组学检测有望发现淋巴瘤早期诊断的血清学“特异性标志蛋白”、不同类型淋巴瘤的“差异蛋白”以及难治性DLBCL的“标志蛋白”,从而早诊断、早治疗并找到难治淋巴瘤的“难治蛋白”进行“靶向治疗”。更多还原

【Abstract】 Objective: To use surface enhanced laser desorption/ionization-time of flight-mass spectrometry ( SELDI-TOF-MS )ProteinChip technology research pathology classification and prognosis of B-cell non-Hodgkin lymphoma To test differentially expressed protein markers of blood serum from B-cell non-Hodgkin lymphoma ( B-cell NHL ),between diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma(FL) patients using surface enhanced laser desorption/ionization-time of flight-mass spectrometry ( SELDI-TOF-MS ) ProteinChip technology. Further, to test differentially expressed protein markers of B-cell NHL patients after chemical therapy using SELDI-TOF-MS ProteinChip technology in order to discuss clinical means of these differentially expressed proteins. Methods: Differentially expressed protein markers of blood serum between 54 B-cell NHL patients and 27 healthy volunteers were analyzing by using SELDI-TOF-MS WCX-2 PorteinChip. Meanwhile differentially expressed protein markers relative to pathology classification - between 23 DLBCL patients and 12 FL patients - were screened; and differentially expressed protein markers which affect prognosis of 23 DLBCL patients were screened. Results: There were five specific marker proteins between 54 B-cell NHL patients and 27 healthy volunteers. Their relative molecular weights were 7974Da, 15938Da, 3398Da, 8564Da, and 8692Da. The protein markers of relative molecular weights 7974Da and 15938Da were high level in patients and the protein markers of relative molecular weights 3398Da, 8564Da and 8692Da were low level in patients. Through efficiently chemical therapy using BACOP, the specific marker proteins which were high level before chemical therapy appeared low level but the specific marker proteins which were low level before chemical therapy appeared high level .There were three differentially expressed protein markers of which relative molecular weights were 3379Da, 3976Da, 4302Da between 23 DLBCL patients and 12 FL patients. They were high level in 23 DLBCL patients and were low level in 12 FL patients. Differentially expressed protein markers of which relative molecular weights were 4795Da and 4998Da affect prognosis of DLBCL patients .The prognosis of DLBCL patients who highly expressed relative molecular weights 4795Da and 4998Da were better. Conclusion: B-cell NHL patients specific marker proteins were 7974Da, 15938Da, 3398Da, 8564Da, and 8692Da. 7974Da and 15938Da were high level in patients and the protein markers of 3398Da, 8564Da and 8692Da were low level in patients. 3379Da, 3976Da, 4302Da protein were high level in 23 DLBCL patients and were low level in 12 FL patients. 4795Da and 4998Da of high level in DLBCL patients’ prognosis were better.SELDI-TOF-MS ProteinChip technology is a quick easy and convenient, and high-throughput analyzing method which can screen several relatively specific protein markers from the blood serum of patients to diagnose B-cell NHL .The relatively specific pr更多还原