重组MUC1多肽及MUC1-MBP融合蛋白的纯化及抗肿瘤作用比较研究

发布时间：2018-01-13 02:17

本文关键词：重组MUC1多肽及MUC1-MBP融合蛋白的纯化及抗肿瘤作用比较研究　出处：《吉林大学》2012年硕士论文　论文类型：学位论文

更多相关文章： MUC1 MUC1-MBP MBP BCG 抗肿瘤

【摘要】：MUC1是粘蛋白家族中的一员，主要分布在腺上皮细胞及其来源的肿瘤细胞表面，因其在正常组织和肿瘤组织的差异使其成为肿瘤疫苗研究的有效靶点。以MUC1为基础的疫苗已经快速进入人类的临床试验，并且表现出良好的疗效。本研究组成功制备出高纯度的MUC1-MBP融合蛋白，初步研究表明，MUC1-MBP蛋白联合BCG免疫在小鼠体内能预防并抑制肺癌转移及实体瘤的生长，，还能明显抑制小鼠皮下人乳腺癌的生长，但对疫苗的各个组分MUC1、MBP、BCG的具体作用还没有进行深入的探讨。本课题的研究目的是在研究组前期研究工作的基础之上进一步比较重组MUC1多肽及MUC1-MBP融合蛋白的抗肿瘤作用，分析MUC1多肽、MBP蛋白、BCG在免疫中所起的作用。本研究通过IPTG诱导MUC1-MBP、MBP及MUC1-GST蛋白表达，采用Amylose resin亲和层析制备高纯度MUC1-MBP融合蛋白及MBP蛋白。用Glutathione Sepherose4B纯化MUC1-GST，采用凝血酶裂解MUC1-GST，获得重组MUC1多肽。用纯化的蛋白在动物肿瘤模型上进行了研究，研究结果表明，MUC1-MBP联合BCG具有显著的抗肿瘤作用，而MUC1多肽联合BCG没有明显的抗肿瘤作用，MUC1-MBP联合BCG作为抗肿瘤疫苗其中的三个组分MUC1、MBP、BCG缺一不可。本研究结果进一步提示了MUC1-MBP联合BCG作为抗肿瘤疫苗的开发价值。
[Abstract]:MUC1 is a member of mucin family, mainly distributed on the surface of glandular epithelial cells and tumor cells derived from them. Because of its difference in normal tissue and tumor tissue, it has become an effective target of tumor vaccine research. The vaccine based on MUC1 has been rapidly introduced into human clinical trials. The high purity MUC1-MBP fusion protein was successfully prepared in this study. MUC1-MBP protein combined with BCG immunized mice can prevent and inhibit the metastasis of lung cancer and the growth of solid tumor, but also can significantly inhibit the growth of subcutaneous human breast cancer in mice. However, MBP was used as a component of the vaccine. The purpose of this study is to compare the recombinant MUC1 peptide and MUC1-MBP fusion protein on the basis of the previous research work of the study group. Antitumor effect. To analyze the role of MUC1 protein in immune system. In this study, the expression of MBP and MUC1-GST protein in MUC1-MBP was induced by IPTG. High purity MUC1-MBP fusion protein and MBP protein were prepared by Amylose resin affinity chromatography. Glutathione was used to prepare the fusion protein. MUC1-GST was purified by Sepherose4B. The recombinant MUC1 peptide was obtained by thrombin cleavage of MUC1-GST.The purified protein was used to study the animal tumor model. MUC1-MBP combined with BCG had significant anti-tumor effect, while MUC1 polypeptide combined with BCG had no obvious anti-tumor effect. MUC1-MBP and BCG were used as three components of MUC1MBP. The results of this study further suggest the value of MUC1-MBP combined with BCG as an anti-tumor vaccine.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R392.9

【参考文献】