激活素A的真核表达以及中药抗肿瘤激活素A相关机制的初步探讨

发布时间：2018-04-28 23:19

本文选题：ActivinA + 真核表达　；参考：《东北师范大学》2008年硕士论文

【摘要】： 激活素是1986年从猪卵泡液中分离的,是由2个β亚单位借二硫键构成的二聚体糖蛋白。激活素β亚基具有典型的TGF-β超家族结构特征,故激活素属于TGF-β超家族成员。激活素根据其亚单位的结合形式不同,分别形成激活素A、激活素B、激活素AB三种分子结构,三种分子结构的激活素具有类似的生物学活性。它是一种多功能的因子,具有组织特异性,在各种组织形成、细胞生长、分化、发育和成熟过程中起到关键性调节作用,也是机体多种生理和病理过程的重要调节因子。真核表达激活素A对于研究其信号传导通路以及研究机体的许多生理和病理过程有重要意义,也为很多与激活素相关的恶性肿瘤的研究与治疗提供了必要的物质基础。本研究将pcDNA3- ActivinβA重组质粒转入CHO细胞,并以转染pcDNA3质粒做对照组。然后在G418选择压力下持续筛选3周,从而获得稳定表达激活素A的细胞株。报告基因检测及免疫印迹分析结果均显示稳定表达激活素A的细胞株构建成功。激活素能抑制上皮细胞增殖,被认为是抑癌基因。激活素信号通路的紊乱可使细胞逃避激活素介导的生长抑制效应,导致多种肿瘤发生。本课题组以往的研究结果显示,睾丸特异性蛋白50(TSP50)具有促进肿瘤发生的作用,而其主要机制是通过抑制激活素信号而使细胞发生恶性增殖,那么,具有抑制TSP50表达的中药单体TI13、TI33、TI176及TI179所具有的抗肿瘤作用是否和激活素信号通路有关?我们用激活素应答性报告载体CAGA-lux转染293T细胞,并分别加入这些中药单体,检测这些单体对激活素信号的影响。结果显示,中药单体TI13、TI176及TI179均可明显增强激活素的信号。本研究为进一步研究激活素在肿瘤发生中的作用及机制奠定了实验基础。
[Abstract]:Activin was isolated from porcine follicular fluid in 1986 and is a dimer glycoprotein composed of two 尾 subunits by disulfide bond. Activin 尾 subunit has typical structural characteristics of TGF- 尾 superfamily, so activin belongs to TGF- 尾 superfamily. According to the different binding forms of activin, activin A, activin B, activin AB are formed respectively. The activin of three molecular structures has similar biological activity. It is a multifunctional factor with tissue specificity. It plays a key role in the process of tissue formation, cell growth, differentiation, development and maturation. It is also an important regulatory factor in many physiological and pathological processes. Eukaryotic expression of activin A plays an important role in the study of its signal transduction pathway and in many physiological and pathological processes of the body. It also provides a necessary material basis for the study and treatment of many malignant tumors associated with activin. In this study, pcDNA3-Activin 尾 A recombinant plasmid was transfected into CHO cells and transfected with pcDNA3 plasmid as control group. Then the cell lines expressing activin A stably were obtained after 3 weeks of continuous screening under G418 selection pressure. The results of reporter gene detection and Western blotting analysis showed that the cell line expressing activin A stably was successfully constructed. Activin inhibits the proliferation of epithelial cells and is thought to be a tumor suppressor gene. The disturbance of activin signaling pathway causes cells to escape activin-mediated growth inhibition, leading to multiple tumorigenesis. Our previous studies have shown that testicular specific protein 50 (TSP50) can promote tumorigenesis, and its main mechanism is to induce malignant proliferation of cells by inhibiting activin signal. Does the anti-tumor effect of TI13, TI33, TI176 and TI179, which inhibits the expression of TSP50, be related to the activin signaling pathway? We transfected 293T cells with activin responsive report vector (CAGA-lux) and added these Chinese medicine monomers to detect the effect of these monomers on activin signal. The results showed that TI13 TI176 and TI179 could significantly enhance the signal of activin. This study provides an experimental basis for the further study of the role and mechanism of activin in tumorigenesis.
【学位授予单位】：东北师范大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R341;R285.5

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