本文選題：癌基因　切入點(diǎn)：胃癌　出處：《上海交通大學(xué)》2015年博士論文　論文類型：學(xué)位論文

【摘要】：目前,胃癌在世界范圍內(nèi)仍然是威脅人類健康的一大病癥,尤其在發(fā)展中國(guó)家該種情況尤為明顯。尋找能夠用于胃癌早期診斷、治療的分子靶標(biāo)結(jié)合臨床手術(shù)能夠明顯改善治療效果。本課題主要從以下三部分,詳細(xì)闡述TRIM59在胃癌中的表達(dá)情況,進(jìn)一步探討了TRIM59參與調(diào)控胃癌發(fā)生發(fā)展的生物學(xué)分子機(jī)制。有望將TRIM59作為一種嶄新的分子靶標(biāo)為臨床診斷治療提供一定的幫助。第一部分通過(guò)對(duì)多個(gè)數(shù)據(jù)庫(kù)進(jìn)行生物信息學(xué)檢索分析發(fā)現(xiàn),在諸多癌基因中位于3號(hào)染色體的TRIM家族成員TRIM59在胃癌中顯著高表達(dá),并且TRIM59的表達(dá)水平與病人的預(yù)后狀況有顯著的相關(guān)性。我們進(jìn)一步通過(guò)對(duì)臨床樣本、組織芯片和胃癌細(xì)胞系中TRIM59的mRNA和蛋白表達(dá)水平檢測(cè)發(fā)現(xiàn),TRIM59在病人的胃癌組織中的表達(dá)水平高于在癌旁組織中的表達(dá)水平,胃癌細(xì)胞系中的TRIM59表達(dá)水平也明顯高于胃粘膜正常上皮細(xì)胞(GES-1)。第二部分通過(guò)體外、體內(nèi)功能性實(shí)驗(yàn)發(fā)現(xiàn),TRIM59促進(jìn)腫瘤細(xì)胞的增殖、克隆形成和遷移以及裸鼠體內(nèi)成瘤。我們通過(guò)RT-PCR和western blot功能機(jī)制驗(yàn)證發(fā)現(xiàn),干擾TRIM59基因的表達(dá),細(xì)胞的惡性程度降低,相應(yīng)的調(diào)控腫瘤惡性增殖轉(zhuǎn)移相關(guān)的信號(hào)通路的活性也有所降低。此外,通過(guò)對(duì)TRIM59和p53調(diào)控的信號(hào)通路中主要成分的表達(dá)分析發(fā)現(xiàn)兩者之間具有明顯的負(fù)相關(guān)性。進(jìn)一步對(duì)臨床樣本的相關(guān)因子檢測(cè)發(fā)現(xiàn),TRIM59與p53蛋白及下游的調(diào)控分子的表達(dá)水平也具有明顯的負(fù)相關(guān)性。第三部分通過(guò)免疫共沉淀、蛋白質(zhì)半衰期檢測(cè)等功能實(shí)驗(yàn)驗(yàn)證發(fā)現(xiàn),TRIM59可以與抑癌基因p53蛋白相互結(jié)合,促進(jìn)p53由蛋白酶體依賴的泛素化降解過(guò)程,進(jìn)而調(diào)節(jié)p53的蛋白穩(wěn)定性,影響其下游基因的表達(dá)最終促進(jìn)了腫瘤的發(fā)生、發(fā)展。綜上所述,我們發(fā)現(xiàn)一個(gè)新的胃癌標(biāo)志物TRIM59,通過(guò)擬合TRIM59表達(dá)水平與病人臨床癥狀相關(guān)性發(fā)現(xiàn),TRIM59可以作為鑒定胃癌發(fā)生發(fā)展的診斷分子,有望為臨床診斷提供一定的參考意義。其次,我們通過(guò)功能驗(yàn)證實(shí)驗(yàn)發(fā)現(xiàn),TRIM59可以激活腫瘤發(fā)展相關(guān)信號(hào)通路,促進(jìn)癌細(xì)胞的增殖、克隆形成、遷移和裸鼠體內(nèi)成瘤。最后,我們發(fā)現(xiàn)TRIM59是通過(guò)調(diào)節(jié)p53的泛素化水平,影響p53蛋白的穩(wěn)定性,進(jìn)而影響p53蛋白的轉(zhuǎn)錄活性促使腫瘤發(fā)生發(fā)展。這些研究可以為臨床診斷治療提供一定的依據(jù)。
[Abstract]:At present, gastric cancer remains a major threat to human health worldwide, especially in developing countries. Molecular target therapy combined with clinical surgery can significantly improve the therapeutic effect. In this paper, the expression of TRIM59 in gastric cancer is described in detail in the following three parts. The biological molecular mechanism of TRIM59 involved in the regulation of carcinogenesis and development of gastric cancer is further discussed. It is expected that TRIM59 will be used as a new molecular target for clinical diagnosis and treatment. The analysis of bioinformatics retrieval found that, TRIM59, a member of the TRIM family located on chromosome 3 of many oncogenes, was significantly overexpressed in gastric cancer, and the expression level of TRIM59 was significantly correlated with the prognosis of patients. The expression level of TRIM59 mRNA and protein in tissue microarray and gastric cancer cell line was higher than that in paracancerous tissue. The expression level of TRIM59 in gastric cancer cell line was also significantly higher than that in normal gastric mucosal epithelial cell line. In the second part, it was found that TRIM59 promoted the proliferation of tumor cells through in vitro functional experiments. Clone formation and migration as well as tumorigenesis in nude mice. We have found that interfering with the expression of TRIM59 gene reduces the malignancy of cells by verifying the functional mechanism of RT-PCR and western blot. The activity of signal pathway associated with malignant proliferation and metastasis is also decreased. By analyzing the expression of the main components in the signal pathway regulated by TRIM59 and p53, we found that there was a significant negative correlation between them. Further, we found that TRIM59, p53 protein and downstream regulatory molecules were detected by the correlation factor detection of clinical samples. The expression level was also negatively correlated. The third part was co-immunoprecipitation. Functional experiments, such as protein half-life detection, showed that TRIM59 could bind to p53 protein, promote the proteasome dependent Ubiquitin degradation process, and regulate the protein stability of p53. Affecting the expression of genes downstream of the tumor ultimately promotes the development of the tumor. We found a new marker of gastric cancer, TRIM59. by fitting the expression level of TRIM59 with the clinical symptoms of patients, we found that TRIM59 can be used as a diagnostic molecule to identify the occurrence and development of gastric cancer, and it is expected to provide a certain reference for clinical diagnosis. We found that TRIM59 can activate the signal pathway associated with tumor development, promote cancer cell proliferation, clone formation, migration and tumorigenesis in nude mice. Finally, we found that TRIM59 regulates the level of p53 ubiquification. The stability of p53 protein is affected, and the transcription activity of p53 protein is affected to promote the tumorigenesis and development. These studies can provide some evidences for clinical diagnosis and treatment.
【學(xué)位授予單位】：上海交通大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2015
【分類號(hào)】：R735.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

1 Pei-Fei Li;Sheng-Can Chen;Tian Xia;Xiao-Ming Jiang;Yong-Fu Shao;Bing-Xiu Xiao;Jun-Ming Guo;;Non-coding RNAs and gastric cancer[J];World Journal of Gastroenterology;2014年18期

2 Wanqing Chen;Rongshou Zheng;Siwei Zhang;Ping Zhao;Hongmei Zeng;Xiaonong Zou;Jie He;;Annual report on status of cancer in China,2010[J];Chinese Journal of Cancer Research;2014年01期

3 KatherineDCrew;AlfredINeugut;;Epidemiology of gastric cancer[J];World Journal of Gastroenterology;2006年03期

4 ;Activation of STAT3 signaling in human stomach adenocarcinoma drug-resistant cell line and its relationship with expression of vascular endothelial growth factor[J];World Journal of Gastroenterology;2005年06期

，

本文編號(hào)：1632775