Maspin基因表达调控的初步研究

发布时间：2018-07-05 07:10

本文选题：Maspin基因 + 启动区　；参考：《山东大学》2005年博士论文

【摘要】：前列腺癌是威胁男性生命的主要疾病之一,关于其侵袭和转移的分子机制需要更深入的研究。Maspin是丝氨酸蛋白酶抑制剂超家族中的一员,在前列腺癌的侵袭与转移中起着非常重要的作用。Maspin表达于正常的哺乳动物上皮细胞中,在原发性前列腺癌细胞中表达下降,在转移性前列腺癌中表达缺失。对maspin基因功能的研究表明:Maspin在体外可以抑制人前列腺癌细胞的侵袭和移动,体内可以抑制裸鼠前列腺癌细胞的生长与转移。而且,Maspin是一有效的血管生成的抑制剂。把前列腺癌细胞LNCaP植入裸鼠的皮下,再用外源性的重组Maspin治疗裸鼠,发现Maspin可以抑制肿瘤细胞的生长和血管的生成。Maspin不仅在侵袭和转移水平抑制前列腺癌的发展,还可以增强肿瘤细胞的凋亡。无论是体外还是体内,内皮细胞中maspin基因的过表达都可以诱导内皮细胞的凋亡,其诱导作用可以被Caspase的抑制剂阻断。Maspin的表达谱及其生物学功能都说明,Maspin可以发展成为一种有效的治疗前列腺肿瘤的靶点。因此,我们有必要在前列腺癌细胞中研究maspin基因的表达调控。 Maspin基因定位于人染色体18q21.3,编码约3.0kb的mRNA转录体,然后翻译成42kDa的蛋白质。Maspin主要存在于多种器官的上皮细胞的细胞膜和细胞质中,细胞核中也有一定的表达。Maspin的cDNA是从正常人的上皮细胞中分离获得,包括2584个核苷酸。对正常细胞和肿瘤细胞中maspin基因的序列分析发现:在肿瘤细胞中maspin基因的序列和结构并没有改变,推测maspin基因在肿瘤细胞中表达下降或缺失并不是基因突变引起的。本研究中我们克隆了含有(-764～+83)的847bp的maspin启动子的报告基因载体,并利用核酸缺失突变技术,报告基因分析和电泳迁移率改变分析(EMSA),在maspin基因启动区鉴定了影响其表达的两个
[Abstract]:Prostate cancer is one of the major life-threatening diseases in men. The molecular mechanism of its invasion and metastasis needs further study. Maspin is a member of the superfamily of serine protease inhibitors. Maspin is expressed in normal mammalian epithelial cells, decreased in primary prostate cancer cells, and absent in metastatic prostate cancer. Studies on the function of maspin gene showed that in vitro, WMaspin could inhibit the invasion and migration of human prostate cancer cells, and inhibit the growth and metastasis of nude mice prostate cancer cells in vivo. Moreover, Maspin is an effective inhibitor of angiogenesis. LNCaP was implanted subcutaneously into nude mice and then treated with exogenous recombinant Maspin. It was found that Maspin could inhibit the growth of tumor cells and angiogenesis. Maspin not only inhibited the development of prostate cancer at the level of invasion and metastasis. It can also enhance the apoptosis of tumor cells. In vitro and in vivo, overexpression of maspin gene in endothelial cells can induce apoptosis of endothelial cells. The expression profile of Maspin and its biological function can be blocked by the inhibitor of Caspase, which indicates that Maspin can be an effective target for the treatment of prostate cancer. Therefore, it is necessary to study the expression and regulation of maspin gene in prostate cancer cells. Maspin gene is located on human chromosome 18q21.3 and encodes about 3.0kb transcription. Then the 42kDa protein. Maspin mainly exists in the cell membrane and cytoplasm of epithelial cells in many organs, and the cDNA of Maspin is isolated from normal human epithelial cells, including 2 584 nucleotides. The sequence analysis of maspin gene in normal cells and tumor cells showed that the sequence and structure of maspin gene did not change in tumor cells. It is speculated that the decrease or deletion of maspin gene expression in tumor cells is not caused by gene mutation. In this study, we cloned the reporter gene vector of maspin promoter containing (-764 ~ 83) 847bp, and identified two genes affecting its expression in the promoter region of maspin gene by using nucleic acid deletion mutation, reporter gene analysis and electrophoretic mobility shift analysis (EMSA).
【学位授予单位】：山东大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R346

【相似文献】