本文選題：沃納綜合癥(WS)　切入點(diǎn)：小鼠胚胎成纖維細(xì)胞　出處：《昆明理工大學(xué)》2016年碩士論文

【摘要】：胃癌在世界范圍內(nèi)是最常見的惡性腫瘤之一,其發(fā)病死亡率高,早期診斷率低。根據(jù)世界衛(wèi)生組織(WHO)公布的全球統(tǒng)計(jì)報(bào)告,世界胃癌年發(fā)病率為13.86%/10萬人,僅次于肺癌發(fā)病率而位居第二。我國(guó)是胃癌的高發(fā)區(qū),每年新發(fā)現(xiàn)40萬胃癌患者,占世界胃癌發(fā)病人數(shù)的42%。惡性腫瘤是嚴(yán)重威脅人類生命與健康的一大類疾病,據(jù)WHO報(bào)道,癌癥將取代心血管疾病成為世界死亡人數(shù)最多的疾病。亞洲及南美洲國(guó)家的胃癌發(fā)病率遠(yuǎn)高于美國(guó)及其他西歐國(guó)家。引起胃癌發(fā)病的機(jī)理仍不是非常清楚,對(duì)于胃癌的基礎(chǔ)研究目前越來越受到各國(guó)研究人員的重視。由于胃癌的高發(fā)年齡主要集中在中老年人群中,因此構(gòu)建一個(gè)衰老動(dòng)物模型來研究胃粘膜細(xì)胞的生理及病理變化將為基礎(chǔ)研究提供一個(gè)獨(dú)特的思路,對(duì)模擬衰老與胃部病變乃至胃癌的發(fā)生都將有所助益。早老綜合癥(Werner Syndrome,WS)是一種罕見的常染色體隱性遺傳病,大約一百萬人中會(huì)有一人罹患此病,該病也被視為是研究衰老的理想模型。為了在小鼠模型中模擬人類的早老綜合癥,我們通過雙敲除Wm基因和端粒酶RNA組分來實(shí)現(xiàn)。在臨床研究中發(fā)現(xiàn),胃癌細(xì)胞中周期阻滯因子p21(Cdknla)表達(dá)異常。已有研究表明由端粒功能異常引起的DNA損傷信號(hào)中p21起到了很重要的作用。然而,p21與衰老相關(guān)的凋亡以及與胃部組織細(xì)胞的增殖活性之間的關(guān)系仍不明確。在本研究中,我們將較晚代端粒RNA組分敲除的小鼠(G2或G3mTerc-/-)和Wm基因敲除以及p2l基因敲除的小鼠進(jìn)行雜交,得到G2或G3mTerc-/-Wrn-/-p21-/-小鼠。最終我們得到子代為G3mTerc-/-Wrn-/-和G3mTerc-/-wrn-/-p21-/-小鼠成纖維細(xì)胞(MEFs)以及G4mTerc-/-Wrn-/-和G4mTerc-/-Wrn-/-p21-/-小鼠并通過western blot技術(shù)分析檢測(cè)MEFs凋亡相關(guān)蛋白的表達(dá)情況。G4 mTerc-/-Wrn-/-p21-/-小鼠出現(xiàn)明顯的體格和活動(dòng)力下降。運(yùn)用流式細(xì)胞儀檢測(cè)分析線粒體膜電位以及細(xì)胞凋亡情況,使用熒光免疫分析技術(shù)分析DNA損傷情況。結(jié)果顯示,p21的功能缺失增強(qiáng)G3 mTerc-/-Wrn-/-MEFs凋亡增強(qiáng)以及DNA損傷增加。此外,通過SA-β-Gal染色法探究G4 mTerc-/-Wrn-/-p21-/-小鼠胃部組織的衰老情況。我們發(fā)現(xiàn)p21的功能缺失不僅沒有緩解胃部組織細(xì)胞的衰老情況,而且能夠加速小鼠胃部組織的衰老進(jìn)程。另外,我們用BrdU摻入法評(píng)估G4mTerc-/-Wrn-/-p21-/-小鼠胃部組織細(xì)胞的增殖率。有意思的是,結(jié)果顯示G4mTerc-/-Wrn-/-p21-/-小鼠胃部組織細(xì)胞的增殖效率顯著提高。
[Abstract]:Gastric cancer is one of the most common malignant tumors in the world.According to the World Health Organization (WHO) global statistics report, the annual incidence of gastric cancer in the world is 138.6% of 100,000 people, second only to the incidence of lung cancer.China is a high incidence area of gastric cancer. 400000 new gastric cancer patients are found every year, accounting for 42 percent of the world gastric cancer.Malignant tumor is a serious threat to human life and health, according to WHO, cancer will replace cardiovascular disease to become the world's largest number of deaths.The incidence of gastric cancer in Asia and South America is much higher than in the United States and other Western European countries.The pathogenesis of gastric cancer is still unclear, and the basic research of gastric cancer has been paid more and more attention by researchers all over the world.Since the high incidence age of gastric cancer is mainly concentrated in the middle-aged and elderly population, the establishment of an aging animal model to study the physiological and pathological changes of gastric mucosal cells will provide a unique way of thinking for basic research.It will be helpful to simulate senescence, gastric lesion and even gastric cancer.Werner SyndromeWS), a rare autosomal recessive disease that affects about one in 1 million people, is also considered an ideal model for the study of aging.In order to mimic human premature syndrome in mouse models, we performed double knockout of the Wm gene and telomerase RNA components.In clinical studies, abnormal expression of cell cycle arrest factor p21 CDknlawas found in gastric cancer cells.It has been shown that p21 plays an important role in the signal of DNA damage caused by abnormal telomere function.However, the relationship between p21 and aging-related apoptosis and gastric tissue cell proliferation is still unclear.鏈，

本文編號(hào)：1705523