黑色素瘤中硫氧还蛋白促进Treg细胞浸润的效应与机制研究

发布时间：2018-04-11 09:26

本文选题：黑色素瘤 + 氧化还原调节　；参考：《暨南大学》2016年博士论文

【摘要】：已有研究表明肿瘤的发生发展与氧化还原平衡密切相关,近期的研究发现抗氧化剂对肿瘤免疫具有复杂的调控作用。硫氧还蛋白(Trx)是一种分布广泛的抗氧化酶,能够还原蛋白质的半胱氨酸残基,使得巯基和二硫键之间相互转换。调节性T细胞(Treg),也称抑制性T细胞(Suppressor T cell),在体外和体内都显示出很强的免疫抑制活性。虽然Treg与恶性肿瘤免疫抑制的关系还在研究之中,但小鼠体内抑制Treg的研究结果表明机体抗肿瘤免疫能力得到了显著提升。本项目重点研究硫氧还蛋白在黑色素瘤中对Treg的调节作用及对肿瘤免疫应答的影响。主要内容分为三个部分:第一部分收集黑色素瘤肿瘤样品,通过免疫组化方法确定Trx在肿瘤样品中的表达变化;通过流式细胞分选技术,检测Treg细胞在黑色素瘤组织中的浸润情况;第二部分通过构建Trx过表达和敲低的黑色素瘤细胞系,在荷瘤小鼠体内研究Trx对Treg细胞浸润的影响;第三部分使用重组的Trx蛋白和Trx中和抗体,研究Trx促进Treg细胞浸润的机制。实验结果如下:1.Trx在人类黑色素瘤组织中高表达,并与肿瘤的分级分期呈正相关性;2.体外培养的黑素瘤细胞可以分泌大量的Trx蛋白;3.Trx可以促进Treg细胞在黑色素瘤中的浸润,从而提高肿瘤的免疫抑制能力;4.在体外细胞和体内荷瘤鼠实验中,Trx可以提高Treg的迁移能力;5.在体外细胞和体内荷瘤鼠实验中,Trx可以促进效应性T细胞向Treg细胞转化;6.在体外细胞和体内荷瘤鼠实验中,Trx可以提高Treg细胞的抗凋亡能力;7.Trx抗体可以抑制Treg细胞在肿瘤局部的浸润,增强抗肿瘤免疫应答;8.Trx抗体可以降低黑色素瘤的生长速率和转移速率。综上所述,本研究明确了Trx在黑色素瘤中对Treg浸润的调节作用,初步分析了Trx促进Treg细胞浸润的机理,明确了Trx抗体在黑色素瘤免疫应答过程中潜在应用价值,为黑色素瘤的治疗提供了新的潜在药物靶点。
[Abstract]:Studies have shown that the occurrence and development of tumor and oxidation reduction equilibrium is closely related to the recent study found that antioxidants play a role in the regulation of tumor immune complex. Thioredoxin (Trx) is a kind of widely distribution of antioxidant enzymes, which reduced cysteine residues of proteins, the conversion between thiol and regulatory two disulfide bonds. T cells (Treg), also known as suppressor T cells (Suppressor T cell), both in vitro and in vivo showed strong immunosuppressive activity. Although the relationship between Treg and malignant tumor is still on immunosuppression, but research results show that the inhibition of Treg in vivo anti-tumor immune ability has been significantly improved. The project focuses on the thioredoxin regulatory role of Treg in melanoma and effect on tumor immune response. The main content is divided into three parts: the first part of the collection of melanoma tumor Tumor samples, determine the expression of Trx in tumor samples by immunohistochemical method; through flow cytometry, the infiltration of Treg cells were detected in melanoma tissues; the second part through the construction of Trx overexpression and knockdown melanoma cell lines, the effect of Trx in tumor bearing mice infiltration the third part of Treg cells; the use of recombinant Trx protein and Trx neutralizing antibody, study mechanism of Trx promoting Treg cell infiltration. The experimental results are as follows: the high expression of 1.Trx in human melanoma tissues, and grading and staging of tumor Cheng Zhengxiang; 2. cultured melanoma cells can secrete Trx protein; 3.Trx can promote the invasion of Treg cells in melanoma, thereby enhancing tumor immune suppression ability; 4. in vitro and in vivo tumor bearing rats, Trx can enhance the migration ability of Treg in vitro; 5. fine Cellular and in vivo tumor bearing rats, Trx can be transformed to Treg cells promote effector T cells in vitro and in vivo; 6. tumor bearing rats, Trx can improve the anti apoptosis ability of Treg cells; 7.Trx antibody can inhibit Treg cells in tumor infiltration, enhanced anti-tumor immune response; 8.Trx antibody to reduce the growth rate and the transfer rate of melanoma. In summary, this study defines Trx in melanoma on the regulation of Treg invasion, preliminary analysis of the mechanism of Trx promoting Treg cell infiltration, the potential application value of Trx antibody in melanoma immune response process, provide potential drug targets the new treatment for melanoma.

【学位授予单位】：暨南大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R739.5

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