本文選題：ActivinA + 真核表達(dá)��； 參考：《東北師范大學(xué)》2008年碩士論文

【摘要】： 激活素是1986年從豬卵泡液中分離的,是由2個β亞單位借二硫鍵構(gòu)成的二聚體糖蛋白。激活素β亞基具有典型的TGF-β超家族結(jié)構(gòu)特征,故激活素屬于TGF-β超家族成員。激活素根據(jù)其亞單位的結(jié)合形式不同,分別形成激活素A、激活素B、激活素AB三種分子結(jié)構(gòu),三種分子結(jié)構(gòu)的激活素具有類似的生物學(xué)活性。它是一種多功能的因子,具有組織特異性,在各種組織形成、細(xì)胞生長、分化、發(fā)育和成熟過程中起到關(guān)鍵性調(diào)節(jié)作用,也是機(jī)體多種生理和病理過程的重要調(diào)節(jié)因子。真核表達(dá)激活素A對于研究其信號傳導(dǎo)通路以及研究機(jī)體的許多生理和病理過程有重要意義,也為很多與激活素相關(guān)的惡性腫瘤的研究與治療提供了必要的物質(zhì)基礎(chǔ)。本研究將pcDNA3- ActivinβA重組質(zhì)粒轉(zhuǎn)入CHO細(xì)胞,并以轉(zhuǎn)染pcDNA3質(zhì)粒做對照組。然后在G418選擇壓力下持續(xù)篩選3周,從而獲得穩(wěn)定表達(dá)激活素A的細(xì)胞株。報告基因檢測及免疫印跡分析結(jié)果均顯示穩(wěn)定表達(dá)激活素A的細(xì)胞株構(gòu)建成功。 激活素能抑制上皮細(xì)胞增殖,被認(rèn)為是抑癌基因。激活素信號通路的紊亂可使細(xì)胞逃避激活素介導(dǎo)的生長抑制效應(yīng),導(dǎo)致多種腫瘤發(fā)生。本課題組以往的研究結(jié)果顯示,睪丸特異性蛋白50(TSP50)具有促進(jìn)腫瘤發(fā)生的作用,而其主要機(jī)制是通過抑制激活素信號而使細(xì)胞發(fā)生惡性增殖,那么,具有抑制TSP50表達(dá)的中藥單體TI13、TI33、TI176及TI179所具有的抗腫瘤作用是否和激活素信號通路有關(guān)?我們用激活素應(yīng)答性報告載體CAGA-lux轉(zhuǎn)染293T細(xì)胞,并分別加入這些中藥單體,檢測這些單體對激活素信號的影響。結(jié)果顯示,中藥單體TI13、TI176及TI179均可明顯增強(qiáng)激活素的信號。本研究為進(jìn)一步研究激活素在腫瘤發(fā)生中的作用及機(jī)制奠定了實驗基礎(chǔ)。
[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.
【學(xué)位授予單位】：東北師范大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號】：R341;R285.5

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 朱輝,劉國藝,李慶雷,倪江;激活素A對離體培養(yǎng)大鼠卵泡發(fā)育的影響[J];基礎(chǔ)醫(yī)學(xué)與臨床;2001年06期

2 黃新,李定國,陸漢明,王志榮,魏紅山,汪余勤,張晶,徐芹芳;激活素和卵泡抑素mRNA在肝纖維化形成過程中的作用[J];中華肝臟病雜志;2002年02期

，

本文編號：1817352