本文選題：食管癌　切入點：干細胞　出處：《北京協和醫(yī)學院》2017年博士論文　論文類型：學位論文

【摘要】：目前,食管癌占全世界范圍內癌癥死亡率的第六位,在我國位于癌癥死亡率的第四位。食管癌在全世界不同地區(qū)和人種間的發(fā)病率有著很大的不同,因此環(huán)境因素與遺傳因素均與食管癌的發(fā)病相關。食管癌主要包括食管鱗狀細胞癌和食管腺癌兩大病理類型,其中食管鱗狀細胞癌約占食管惡性腫瘤的90%,我國中部地區(qū)及中亞是食管鱗癌的主要流行地區(qū)。在我國食管癌病因學研究領域,上世紀七八十年代通過對河南省林州市食管癌高發(fā)現場深入的病因學研究明確了亞硝胺類物質、霉變食物、微量元素缺乏等是我國食管癌的重要化學病因。這其中,在林縣食管癌高發(fā)現場分離得到的甲基芐基亞硝胺是目前已知最強的誘導大鼠食管癌的化學致癌物,并且其誘導的大鼠食管癌病理形態(tài)與人食管鱗狀細胞癌的發(fā)生時序相平行,因此,甲基芐基亞硝胺主要被用來構建大鼠食管癌動物模型。該動物模型對研究食管癌的發(fā)生、發(fā)展機制及化學預防有著重要意義。對于上皮來源實體瘤的研究如實體瘤內部不同細胞體外克隆形成能力的差異,使得人們對腫瘤異質性有了更深入的認識。腫瘤干細胞理論的提出為解釋腫瘤細胞的異質性提供了一定的理論依據。目前,食管鱗癌干細胞標志物的研究仍處于探索階段,特異程度較高的標志物依然有待進一步尋找。二甲雙胍是目前應用最廣泛的2型糖尿病治療藥物,近期一些流行病學研究顯示二甲雙胍可以降低多種實體瘤的發(fā)生,這其中二甲雙胍降低食管癌發(fā)生的流行病學研究也偶有報道,但二甲雙胍降低食管癌發(fā)生發(fā)展的分子機制卻未見明確報道。本研究中,我們首先構建了兩種NMBzA給藥途徑誘導的大鼠食管癌動物模型并優(yōu)化動物模型,得到了從增生、不典型增生到癌階段的動態(tài)病理標本。免疫組化染色分析了目前被證實的大鼠食管干細胞標志物及人食管鱗癌干細胞標志物等在大鼠食管癌發(fā)生發(fā)展過程中的動態(tài)變化情況。其次,我們利用NMBzA皮下給藥模型進行了二甲雙胍預防食管鱗癌發(fā)生的動物模型研究,證實了二甲雙狐能夠有效抑制NMBzA誘導的大鼠食管癌及其癌前病變的發(fā)生與發(fā)展,并揭示了 AMPK/mTOR信號轉導通路是二甲雙胍抑制食管癌發(fā)生的主要作用靶點。最后,通過體外細胞實驗進一步證實二甲雙胍可以抑制食管上皮永生化細胞與食管癌細胞的增殖能力,使上述細胞的細胞周期阻滯在G0/G1期,并促進細胞凋亡的發(fā)生,上述現象與二甲雙胍抑制食管癌細胞Bcl-2及cyclinD1的表達相關。此外,食管癌細胞的遷移運動能力與平板克隆形成能力及成球性生長能力也顯著受到抑制,這與二甲雙胍影響食管癌細胞中AMPK/mTOR、PI3K/Akt及Stat3信號轉導通路和降低干性相關基因表達相關。綜上所述,我們的研究成功構建了 NMBzA誘導的大鼠食管癌動物模型,為大鼠食管癌發(fā)生發(fā)展過程中各重要基因的突變與表達量的改變以及研究食管鱗癌干細胞的起源問題提供了標本來源。通過體內、體外實驗證實了二甲雙胍預防食管癌發(fā)生發(fā)展的分子機制,為二甲雙胍在臨床中預防及治療食管癌癌前病變提供一定的理論基礎。
[Abstract]:At present, esophageal cancer accounted for worldwide cancer mortality in China in sixth, cancer mortality fourth. The incidence of esophageal cancer in the world in different regions and races are very different, so the incidence of environmental and genetic factors were associated with esophageal cancer esophageal cancer mainly includes two. Pathological types of esophageal squamous cell carcinoma and adenocarcinoma of the esophagus, and esophageal squamous cell carcinoma accounts for about 90% of malignant tumor of esophagus, the central region of China and Central Asia is the major epidemic areas of esophageal cancer. In the field of study on etiology of esophageal cancer in China, the last century in 70s and 80s by the Linzhou city of Henan Province, a high incidence of esophageal cancer etiology research field further clear the nitrosamines, moldy food, lack of trace elements are important chemical etiology of esophageal cancer in China. Among these, the separated methyl in Lin County, a high risk area of esophageal cancer Benzyl nitrosamine induced esophageal cancer in rats is currently the strongest known chemical carcinogens, and the timing of rats induced by esophageal cancer and pathological morphology of human esophageal squamous cell carcinoma are parallel, therefore, METHYLBENZYLNITROSAMINE mainly is used to construct esophageal carcinoma animal model rats. The animal model study on esophageal cancer, has important significance and development mechanism of chemoprevention. For solid tumors of epithelial origin such as differences in tumor cells in vitro in different colony forming ability, which make people have a deeper understanding of tumor heterogeneity. The cell theory provides a theoretical basis for the interpretation of tumor heterogeneity the tumor stem cells. At present, esophageal squamous cell carcinoma stem cell marker research is still in the exploratory stage, the higher degree of specific markers remains to be further search. Metformin is the most Type 2 diabetes wide drug treatment, some recent epidemiological studies suggest that metformin may reduce the occurrence of a variety of solid tumors, which metformin reduces epidemiological study on esophageal cancer have also been reported, but the molecular mechanism of metformin reduces the development of esophageal cancer has not been clearly reported. In this study, and optimize the animal model of esophageal cancer in rats the animal model we first constructed two NMBzA administration induced, obtained from hyperplasia, dysplasia and carcinoma specimens dynamic pathological stage. Immunohistochemical staining analysis confirmed the rat esophageal stem cell markers and human esophageal squamous cell carcinoma stem cell markers have dynamic changes in the development process in esophageal carcinoma rats. Secondly, we use the NMBzA model by subcutaneous administration of metformin on animal model of esophageal carcinoma, syndrome The two armor double fox to the occurrence and development of esophageal carcinoma rats effectively inhibit NMBzA induced and precancerous lesions, and reveals the AMPK/mTOR signal transduction pathway is the main effect of metformin inhibited esophageal cancer target. Finally, through in vitro experiments further confirmed that two metformin can inhibit the proliferation ability of human immortalized esophageal epithelial cells with esophageal cancer cells, block the cell cycle of the cells in the G0/G1 phase, and promotes cell apoptosis, the expression of the above phenomenon with metformin inhibits Bcl-2 and cyclinD1 esophageal cancer cells. In addition, esophageal cancer cell migration and colony forming ability and Ballability growth ability also significantly inhibited. The effects of metformin and AMPK/mTOR in esophageal carcinoma cells, PI3K/Akt and Stat3 signal transduction pathway and reduce the stemness related genes expression. In summary, I Our study successfully constructed the animal model of NMBzA induced rat esophageal cancer, esophageal cancer rat mutated each important gene during the development and expression and the origin of esophageal squamous cell carcinoma stem cells provide a source of specimen. In vivo, in vitro experiments confirmed the molecular mechanism of the occurrence and development of esophageal and metformin for the prevention of cancer, and provide a theoretical basis for the treatment of esophageal precancerous lesions of metformin in clinic.

【學位授予單位】：北京協和醫(yī)學院
【學位級別】：博士
【學位授予年份】：2017
【分類號】：R735.1

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