卵巢上皮癌多药耐药的血清蛋白组学及代谢组学的初步研究

发布时间：2018-02-21 18:30

本文关键词： 卵巢上皮癌多药耐药蛋白组学代谢组学血清　出处：《广西医科大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的使用蛋白组学及代谢组学技术研究卵巢上皮癌多药耐药患者血清中差异表达的蛋白质及代谢化合物，，寻找卵巢上皮癌多药耐药相关的诊断标志物及探索多药耐药的发病机制，进而找到逆转耐药治疗的靶点。方法收集临床上卵巢上皮癌（EOC）铂类耐药患者（PTR）、非耐药患者（PTS）、卵巢良性囊肿（BOC）及正常健康人（NC）四组血清各10例，每组的10例样品混合成一个pool，使用Human14亲和柱去除血清中高丰度蛋白后，收集去除高丰度蛋白后的组分用于蛋白组学的分析。本实验采用基于iTRAQ的蛋白组学技术分析不同分组血清差异蛋白表达谱，然后借助生物信息学方法筛选出有重要价值的蛋白质，采用Western blot及ELISA技术验证其表达的规律；此外，从蛋白组学分析的样本中抽取132例临床血清样本，包括PTR、PTS、BOC及NC四组，利用液质联用正离子模式检测四组血清代谢指纹谱。结果两次iTRAQ实验总共鉴定三百多种蛋白，本研究对四组样本中两两进行对比分析，重点关注PTS及PTR间的差异表达，两组鉴定到62个有统计学意义的差异表达蛋白，使用生物信息学方法对这62个蛋白进行全面分析，并采用Western blot及ELISA技术对部分差异蛋白进行验证，表达趋势与质谱一致。代谢组学共检测到25800个代谢化合物，使用主成分分析（PCA）对数据降维后，并结合临床资料进行分析后得到6个对EOC多药耐药意义重大的差异代谢物。结论差异表达蛋白SERPINA1、FN1、ORM1及6个差异代谢物可对EOC化疗的疗效起到监测作用，可能参与EOC多药耐药的生物过程，是EOC多药耐药潜在的诊断标志物及逆转耐药治疗药物的作用靶点。
[Abstract]:The purpose of using protein and metabolic compounds of proteomics and metabolomics technology of differential expression of epithelial ovarian cancer multidrug resistance in the serum of patients with epithelial ovarian cancer, looking for multidrug resistance related markers and explore the pathogenesis of multidrug resistance, and then find the target for drug resistance reversal treatment.
Methods clinical on epithelial ovarian cancer (EOC) patients with platinum resistant (PTR), non resistant patients (PTS), benign ovarian cyst (BOC) and healthy people (NC) between the four groups in all 10 cases, each of the 10 samples are mixed into a pool, use the Human14 affinity column to remove high abundant proteins the serum collected after removing high abundance proteins after the component is used for the analysis of proteomics. In this experiment, the protein iTRAQ of serum protein expression profile differences in different groups based on, and with the help of screening valuable protein bioinformatics methods, using Western blot and ELISA technology to verify the expression of the law; in addition, from proteomic analysis in samples from 132 cases of clinical serum samples, including PTR, PTS, BOC and NC four groups, combined with positive ion mode detection of serum metabolic fingerprinting using four groups of fluid.
The results of the two experiment iTRAQ a total of more than 300 proteins identified in this study, 22 of four samples were analyzed, focusing on the PTS expression and PTR protein expression in 62, there was a significant difference between the two groups identified using bioinformatics methods to analyze the 62 egg white, and validation of some proteins by Western blot and ELISA technology, expressed the same trend with mass spectrometry. Metabonomics detected a total of 25800 metabolic compounds, using principal component analysis (PCA) to reduce the dimensionality of the data, and combined with clinical data were analyzed after 6 significant differences on EOC multidrug resistant metabolites.
Conclusion differentially expressed proteins SERPINA1, FN1, ORM1 and 6 differential metabolites can play a monitoring role in the efficacy of EOC chemotherapy. They may be involved in the biological process of EOC multidrug resistance. It is a potential diagnostic marker for EOC multidrug resistance and a target for reversing drug-resistant drugs.

【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.31

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