急性淋巴细胞白血病患儿血小板蛋白质组学研究

发布时间：2018-04-30 18:44

本文选题：急性淋巴细胞白血病 + 表面激光解吸电离飞行时间质谱　；参考：《泸州医学院》2013年硕士论文

【摘要】：目的急性淋巴细胞白血病（ALL）是儿童时期最常见的血液系统肿瘤，出血是常见症状及死因之一，引起出血的机制复杂，其中最主要原因是血小板减少。ALL患儿血小板数量减少的同时，可能存在其功能异常。目前关于ALL患儿血小板功能的研究较少，其对出血影响的机制尚不完全清楚。本实验旨在应用血小板蛋白质组学技术分析儿童ALL状态下血小板蛋白质差异表达，并对这些差异表达蛋白质进行进一步的研究和筛选，以期寻找到ALL患儿血小板功能相关的血小板特异性蛋白质，探讨ALL时血小板功能异常的分子机制，以便早期采取积极措施干预ALL患者的出血，从而改善预后。方法应用蛋白质芯片CM10及表面激光解吸电离飞行时间质谱技术（SELDI-TOF-MS）技术获得ALL患儿27例（ALL组）、ALL诱导缓解治疗达到完全缓解患儿25例（ALL-CR1组）及正常对照组27例的整套血小板蛋白质谱，采用Ciphergen公司Biomarker Wizard和BiomarkerPattern System5.0软件对蛋白质谱进行差异蛋白分析，筛选出有显著差异的蛋白质。结果(1)在质荷比2000～20000范围内，ALL组与正常对照组比较血小板质谱中有176个差异表达的蛋白质峰（p＜0.05），高表达有25个，低表达有151个。其中存在稳定表达且变异系数小的蛋白质峰有9个：包括高表达2个，荷质比（M/Z）分别为2496.9和4287.9；低表达7个，荷质比（M/Z）分别为7881.2、4091.3、3149.9、2365.1、9414.1、5252.3、2280.7。(2)ALL组与ALL-CR1组比较血小板质谱中有112个差异表达的蛋白质峰（p＜0.05），其中高表达有15个，低表达有97个，存在稳定表达且变异系数小的蛋白质峰有6个：包括高表达2个，荷质比（M/Z）分别为2496.9和4287.9；低表达4个，荷质比（M/Z）分别为9414.1、7881.2、3149.9、2280.7。(3)ALL-CR1组与正常对照组比较，未出现具有统计学意义的高表达蛋白质峰（p＞0.05），但仍有69个显著低表达的蛋白质峰（p＜0.05）。(4)数据库分析ALL组与正常对照组显著差异的9个蛋白质峰，表达上调的两种蛋白分别为内皮素-1和大内皮素-1，，表达下调的蛋白为血小板第四因子、凝血酶轻链、垂体腺苷酸环化酶多肽27、纤维蛋白原β链和三种未知蛋白，这些蛋白质可能与血小板信号转导、粘附、聚集及活化等功能相关。结论(1) ALL患儿与正常对照组外周血血小板蛋白质组学水平有显著差异；(2)ALL组与ALL-CR1组间血小板蛋白质组学水平也存在显著差异，经诱导缓解治疗达完全缓解后异常表达的蛋白数目明显减少，但与正常比较仍有一部分蛋白呈显著低表达。(3)ALL患儿血小板功能可能存在异常，主要表现为细胞信号转导障碍、聚集和活化能力降低及凝血功能异常等方面；(4)本研究为ALL发病及治疗过程中血小板功能的监测提供了新途径。
[Abstract]:Objective Acute lymphoblastic leukemia (ALL) is the most common hematological tumor in childhood. Hemorrhage is one of the common symptoms and causes of death. The main reason is that the number of thrombocytopenia. All may have abnormal function at the same time. At present, there are few studies on platelet function in children with ALL, and the mechanism of its effect on bleeding is not fully understood. The purpose of this study was to analyze the differential expression of platelet proteins in children with ALL by using platelet proteomics, and to further study and screen these differentially expressed proteins. The aim of this study was to find platelet specific proteins related to platelet function in children with ALL and to explore the molecular mechanism of platelet dysfunction in patients with ALL so as to take active measures to intervene the bleeding in patients with ALL early and improve the prognosis. Methods using CM10 and surface laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS) technique, the complete set of 27 cases of ALL children in all group (25 cases of complete remission group) and 27 cases of normal control group (control group) were obtained by means of protein chip CM10 and surface laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS). Platelet protein profile, Biomarker Wizard and BiomarkerPattern System5.0 software of Ciphergen Company were used to analyze the differential protein profiles and the proteins with significant difference were screened out. Results 1) there were 176 differentially expressed protein peaks (p < 0.05), 25 high expression and 151 low expression in all group compared with normal control group within the range of 2000 ~ 2000.The results showed that there were 176 differentially expressed protein peaks (p < 0.05), 25 high expression and 151 low expression in all group as compared with those in normal control group. Among them, there are 9 protein peaks with stable expression and low coefficient of variation, including 2 high expression, 2496.9 and 4287.9, respectively, and 7 low expression. Compared with ALL-CR1 group, there were 112 differentially expressed protein peaks (p < 0.05) in platelets mass spectrometry (P < 0.05), 15 of which were overexpression and 97 were low expression, compared with the ALL-CR1 group, the ratio of charge to substance was 7881.2% 4091.3N 3149.9N 2365.1N 9414.1n 5252.3N 2280.7.2L all group, compared with ALL-CR1 group, there were 112 differentially expressed protein peaks (p < 0.05), among which 15 had high expression and 97 had low expression. There were 6 protein peaks with stable expression and low coefficient of variation, including 2 high expression, M / Z = 2496.9 and 4287.9, respectively, and 4 low expression, respectively. The ratio of M / Z and M / Z were 9414.1 / 7881.23149.9and 2280.7.3ALL-CR1, respectively, compared with the control group. There were no statistically significant high expression protein peaks (p > 0.05), but there were still 69 significant low expression protein peaks (p < 0.05, P < 0.05) in the ALL group and the control group, which were significantly different from those in the control group (P < 0.05), but there were still 69 protein peaks with significant differences between the ALL group and the normal control group. The up-regulated proteins were endothelin-1 and large endothelin-1. The down-regulated proteins were platelet fourth factor, thrombin light chain, pituitary adenylate cyclase polypeptide 27, fibrinogen 尾 chain and three unknown proteins. These proteins may be related to platelet signal transduction, adhesion, aggregation and activation. Conclusion (1) there is a significant difference in the level of platelet proteomics between children with ALL and normal controls. There is also a significant difference in the level of platelet proteomics between all group and ALL-CR1 group. The number of abnormally expressed proteins after induction remission therapy was significantly decreased, but some of the proteins were significantly lower than those of normal children. The platelet function of all children may be abnormal, mainly manifested as cell signal transduction disorder. This study provides a new approach for monitoring platelet function in the pathogenesis and treatment of ALL.
【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R733.71

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