人肝再生增强因子相互作用蛋白的研究

发布时间：2018-11-25 10:27

【摘要】： 目的:肝再生增强因子(augmenter of liver regeneration, ALR)是近年来发现的一种促肝细胞再生的细胞因子。ALR的生物学作用主要表现在直接刺激肝细胞增殖和再生、对肝脏的保护作用、参与组织器官的形成与发育、免疫调控作用以及促肝癌细胞增殖等效应。尽管有上述重要生物学作用,但ALR究竟如何发挥生物学作用目前仍知之较少。如果能获得与ALR发生相互作用的蛋白,并且鉴定出ALR受体,将有助于深刻理解ALR的生物学功能,特别是为深入了解ALR与肝癌发生、发展的关系奠定基础。本实验拟从人肝癌细胞的cDNA噬菌体表面展示文库中筛选与人肝再生增强因子(human augmenter of liver regeneration, hALR)相互作用的特异噬菌体克隆,通过序列分析及生物学活性检测确认hALR相互作用蛋白,构建原核表达载体,为进一步鉴定ALR受体提供实验基础。方法: 1.以hALR为靶蛋白,从人肝癌细胞cDNA噬菌体展示文库中筛选与hALR相互作用的特异噬菌体克隆;对获得的特异噬菌体克隆cDNA插入片段进行序列测序及生物信息学分析;利用3H-TdR掺入法测定hALR联合阳性噬菌体展示肽及单独阳性噬菌体展示肽对QGY肝癌细胞株的增殖效应。
[Abstract]:Objective: (augmenter of liver regeneration, ALR) is a kind of cytokine found in recent years to promote the regeneration of hepatocytes. The biological effects of ALR are mainly manifested in the direct stimulation of hepatocyte proliferation and regeneration and the protective effect of ALR on the liver. Participate in the formation and development of tissues and organs, immune regulation and promote the proliferation of liver cancer cells. Despite these important biological roles, little is known about how ALR plays its biological role. If we can obtain the protein that interacts with ALR and identify the ALR receptor, it will be helpful to understand the biological function of ALR, especially to lay a foundation for further understanding the relationship between ALR and the occurrence and development of HCC. The aim of this study was to screen specific phage clones interacting with human liver regeneration enhancer (human augmenter of liver regeneration, hALR) from cDNA phage display library of human hepatoma cells. The hALR interacting protein was confirmed by sequence analysis and biological activity detection, and the prokaryotic expression vector was constructed, which provided the experimental basis for further identification of ALR receptor. Methods: 1. Using hALR as the target protein, the specific phage clones interacting with hALR were screened from the cDNA phage display library of human hepatoma cells, and the cDNA inserted fragments were sequenced and analyzed by bioinformatics. The proliferative effects of hALR combined with phage display peptide and single positive phage display peptide on QGY hepatoma cell line were determined by 3H-TdR incorporation.
【学位授予单位】：重庆医科大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R341

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