本文選題：食管鱗狀細胞癌 + 蛋白質(zhì)組學(xué)��； 參考：《新疆醫(yī)科大學(xué)》2015年博士論文

【摘要】：目的:研究食管鱗狀細胞癌差異蛋白的篩選及其臨床意義。方法:(1)通過激光捕獲顯微切割技術(shù)分別從活檢的新鮮腫瘤組織獲取哈薩克族、漢族食管鱗癌細胞和癌旁遠端食管正常上皮細胞的純化細胞。(2)通過雙向電泳聯(lián)合基質(zhì)輔助激光解吸電離飛行時間質(zhì)譜檢測獲得表達差異蛋白。(3)使用蛋白質(zhì)印跡和免疫組化技術(shù)對差異候選蛋白Bmi-1,PAI-1,Cofilin-1和Transgelin進行分析、驗證。(4)對Bmi-1,PAI-1,Cofilin-1,Transgelin與食管鱗狀細胞癌臨床病理參數(shù),如性別、年齡、民族、腫瘤大小、分化程度、病理分型、浸潤深度、淋巴結(jié)轉(zhuǎn)移、臨床分期,及預(yù)后等方面的關(guān)聯(lián),初步探討B(tài)mi-1,PAI-1,Cofilin-1和Transgelin在食管癌發(fā)生發(fā)展中的作用。結(jié)果:(1)激光捕獲顯微切割技術(shù)捕獲了較純凈的哈薩克族和漢族的食管鱗癌細胞及癌旁遠端食管正常上皮細胞。(2)通過雙向電泳聯(lián)合基質(zhì)輔助激光解吸電離飛行時間質(zhì)譜鑒定出43個有意義的差異蛋白,其中在哈薩克族和漢族的食管鱗癌組織表達均上調(diào)的點有25個。在哈薩克族和漢族的食管鱗癌組織均下調(diào)的蛋白點有14個。還有4個蛋白在哈薩克族和漢族食管鱗癌組織中表達不一致。(3)在哈薩克族和漢族的食管鱗癌組織表達均上調(diào)的蛋白中,根據(jù)蛋白得分值,匹配肽段覆蓋率,結(jié)合研究報道較少等因素,從中篩選出四個蛋白質(zhì)Bmi-1,PAI-1,Cofilin-1和Transgelin。Bmi-1,PAI-1,Cofilin-1和Transgelin在哈薩克族、漢族食管鱗癌組織中的表達經(jīng)過蛋白質(zhì)印跡和免疫組化驗證,表達明顯高于食管正常組織。免疫組化結(jié)果:Bmi-1,PAI-1,Cofilin-1和Transgelin在食管鱗癌組織中表達的陽性率為78.75%(P0.05)、76.25%(P0.05)、60.00%(P0.05)、55.00%(P0.05),高于在正常食管組織的表達。(4)分析Bmi-1,PAI-1,Cofilin-1和Transgelin在食管鱗癌與性別、年齡、民族、腫瘤大小、分化程度、病理分型、浸潤深度、淋巴結(jié)轉(zhuǎn)移、臨床分期的表達情況。Bmi-1的表達:在有淋巴結(jié)轉(zhuǎn)移和無淋巴結(jié)轉(zhuǎn)移食管鱗癌患者組織中,Bmi-1的陽性表達率分別為63.0%和86.8%,P0.05,兩組之間差異有統(tǒng)計學(xué)意義。在不同浸潤深度食管鱗癌患者組織中Bmi-1的陽性表達率,腫瘤細胞侵潤食管粘膜或肌層為92.3%,浸潤食管纖維膜及周圍結(jié)構(gòu)為72.2%,P0.05,差異有統(tǒng)計學(xué)意義,結(jié)果表明Bmi-1的表達可能與食管癌組織的浸潤程度有關(guān)。PAI-1的表達:在食管鱗癌患者中,在有淋巴結(jié)轉(zhuǎn)移和無淋巴結(jié)轉(zhuǎn)移食管鱗癌患者組織中,PAI-1的陽性表達率分別為59.3%和84.9%,P0.05,兩組之間差異有統(tǒng)計學(xué)意義,表明PAI-1的表達可能與食管癌組織的淋巴結(jié)轉(zhuǎn)移有關(guān)。腫瘤細胞浸潤食管粘膜或肌層為53.8%,腫瘤浸潤食管纖維膜及周圍結(jié)構(gòu)陽性表達率87.0%,P0.05,差異有統(tǒng)計學(xué)意義,結(jié)果表明PAI-1的表達可能與食管癌組織的浸潤遷移有關(guān)。Cofilin-1的表達:食管鱗癌患者中,在高分化、中分化和低分化食管鱗癌患者組織中Cofilin-1的陽性表達率分別為26.1%、78.3%和70.6%,P0.05,差異有統(tǒng)計學(xué)意義,表明Cofilin-1的表達可能與食管癌組織的分化程度有關(guān),提示表達率高惡性程度高,與預(yù)后相關(guān)。在有淋巴結(jié)轉(zhuǎn)移和無淋巴結(jié)轉(zhuǎn)移食管鱗癌患者組織中,Cofilin-1的陽性表達率分別為88.9%和45.3%,P0.05,兩組之間差異有統(tǒng)計學(xué)意義,表明Cofilin-1的表達與食管癌組織的淋巴結(jié)轉(zhuǎn)移有關(guān)。在臨床分期Ⅰ~Ⅱ期和Ⅲ~Ⅳ期食管鱗癌患者組織中Cofilin-1的陽性表達率分別為37.1%和77.8%,P0.05,兩組之間差異有統(tǒng)計學(xué)意義,表明Cofilin-1的表達可能與食管癌的臨床分期有關(guān)。Transgelin的表達:在高分化、中分化和低分化食管鱗癌患者組織中Transgelin的陽性表達率分別為34.8%、52.2%和70.6%,P0.05,差異有統(tǒng)計學(xué)意義,表明Transgelin的表達可能與食管癌組織的分化程度有關(guān),提示表達率高惡性程度高,與預(yù)后相關(guān)。在有淋巴結(jié)轉(zhuǎn)移和無淋巴結(jié)轉(zhuǎn)移食管鱗癌患者組織中,Transgelin的陽性表達率分別為88.9%和37.7%,P0.05,兩組之間差異有統(tǒng)計學(xué)意義,表明Transgelin的表達與食管癌組織的淋巴結(jié)轉(zhuǎn)移有關(guān)。結(jié)論:(1)本研究提示激光捕獲顯微切割技術(shù)可以有效地解決組織異質(zhì)性的問題,能較好地獲取較為純凈的目的細胞。(2)食管鱗癌和正常食管上皮細胞的差異蛋白質(zhì)圖譜提示食管鱗癌的發(fā)生發(fā)展是多基因參與、多種蛋白質(zhì)功能共同協(xié)同發(fā)揮作用的結(jié)果。(3)蛋白質(zhì)印跡技術(shù)和免疫組化分析Bmi-1,PAI-1,Cofilin-1,Transgelin的驗證結(jié)果與蛋白質(zhì)組學(xué)技術(shù)的分析鑒定結(jié)果相一致。(4)Bmi-l、PAI-1、Cofilin-1和Transgelin這四個蛋白在食管鱗癌組織中表達上調(diào),與食管鱗癌的分化程度、侵潤、淋巴轉(zhuǎn)移有關(guān),在食管鱗狀細胞癌的發(fā)生和發(fā)展中可能起一定作用,并且對食管鱗狀細胞癌惡性程度及其預(yù)后有指導(dǎo)意義,為食管鱗癌分子靶向治療和新的抗癌藥物開發(fā)提供有價值的理論依據(jù)。
[Abstract]:Objective: To study the screening and clinical significance of differential protein in squamous cell carcinoma of the esophagus. Methods: (1) the pure cells of Kazak, Hans squamous cell carcinoma cells and normal epithelial cells near the distal esophagus were obtained by laser capture microdissection. (2) two dimensional electrophoresis combined with matrix assisted laser. The differential proteins were expressed by desorption ionization time of flight mass spectrometry. (3) the difference candidate proteins Bmi-1, PAI-1, Cofilin-1 and Transgelin were analyzed by Western blot and immunohistochemistry. (4) the clinicopathological parameters of Bmi-1, PAI-1, Cofilin-1, Transgelin and esophageal squamous cell carcinoma, such as sex, age, nationality, tumor size, were analyzed. The correlation of differentiation, pathological type, infiltration depth, lymph node metastasis, clinical stage, and prognosis, and the role of Bmi-1, PAI-1, Cofilin-1 and Transgelin in the development of esophageal cancer. Results: (1) laser capture microdissection has captured the purified esophageal squamous cell carcinoma cells and the paracancerous distal end of the Kazak and Han nationality Normal epithelial cells of the esophagus (2) 43 significant differential proteins were identified by two-dimensional electrophoresis combined with matrix assisted laser desorption ionization time of flight mass spectrometry. There were 25 up-regulated points for esophageal squamous cell carcinoma in Kazak and Han people. There were 14 down regulated protein points in the Hazakh and Han squamous cell carcinoma tissues. The expression of 4 proteins in the Kazak and Han esophageal squamous cell carcinoma tissues is inconsistent. (3) in the Hazakh and Han population, the expression of esophageal squamous cell carcinoma is up to up protein, according to the value of the protein score, the coverage rate of the matched peptide segment, and less research reports, four proteins, Bmi-1, PAI-1, Cofilin-1 and Transgelin.Bmi-1, are screened. The expression of PAI-1, Cofilin-1 and Transgelin in the Hazakh and Han esophageal squamous cell carcinoma tissues was verified by Western blot and immunohistochemistry. The expression was significantly higher than that of the normal esophagus. The positive rate of Bmi-1, PAI-1, Cofilin-1 and Transgelin in the tissues of esophageal squamous cell carcinoma was 78.75% (P0.05), 76.25% (P0.05), and 60% (P0.05). 55% (P0.05), higher than the expression of normal esophageal tissue. (4) analysis of the expression of Bmi-1, PAI-1, Cofilin-1 and Transgelin in esophageal squamous cell carcinoma with sex, age, nationality, tumor size, differentiation, pathological classification, infiltration depth, lymph node metastasis, and clinical stage expression of.Bmi-1: lymph node metastasis and lymphatic metastasis of esophageal squamous cell carcinoma. The positive rates of Bmi-1 were 63% and 86.8% in the patients' tissues, respectively, and 86.8%, respectively. The difference between the two groups was statistically significant. The positive rate of Bmi-1 in the tissues of the patients with different infiltrating depth of esophageal squamous cell carcinoma, the tumor cells infiltrating the esophageal mucosa or the myometrium were 92.3%, the infiltrating esophagus membrane and the surrounding structure were 72.2%, and the difference was statistically significant. The results show that the expression of Bmi-1 may be related to the expression of.PAI-1 in the invasive degree of esophageal carcinoma: in the patients with esophageal squamous cell carcinoma, the positive expression rate of PAI-1 is 59.3% and 84.9% in the tissues of the patients with lymph node metastasis and no lymph node metastasis, respectively, and the difference between the two groups is statistically significant, indicating that the expression of PAI-1 can be expressed. It was related to lymph node metastasis of esophageal carcinoma tissue. The tumor cells infiltrated the esophagus mucosa or myometrium 53.8%, the positive expression rate of the tumor infiltrating esophageal fibrous membrane and surrounding structure was 87%, P0.05, the difference was statistically significant. The results showed that the expression of PAI-1 may be related to the expression of.Cofilin-1 in the infiltration and migration of esophageal cancer tissue: the patients with esophageal squamous cell carcinoma The positive expression rates of Cofilin-1 in the tissues of highly differentiated, medium and low differentiated esophageal squamous cell carcinoma were 26.1%, 78.3% and 70.6%, and P0.05, respectively. The difference was statistically significant, indicating that the expression of Cofilin-1 may be related to the degree of differentiation of esophageal cancer tissue, suggesting that the expression rate is high and is associated with the prognosis. There are lymph node metastasis and no The positive rates of Cofilin-1 were 88.9% and 45.3% in the lymph node metastasis of esophageal squamous cell carcinoma, P0.05, respectively, and the difference between the two groups was statistically significant, indicating that the expression of Cofilin-1 was related to the lymph node metastasis of the esophageal carcinoma. The positive expression rate of Cofilin-1 in the tissues of the patients with esophageal squamous cell carcinoma in stage I and II stage and in stage III to IV stage were divided. The difference between 37.1% and 77.8%, P0.05, and two groups was statistically significant, indicating that the expression of Cofilin-1 may be related to the expression of.Transgelin in the clinical stages of esophageal cancer: the positive expression rate of Transgelin in highly differentiated, moderately differentiated and poorly differentiated esophageal squamous cell carcinoma tissues is 34.8%, 52.2% and 70.6%, and P0.05, respectively, with statistical significance. The expression of Transgelin may be related to the degree of differentiation of esophageal cancer tissue, suggesting that the expression rate is high and is associated with the prognosis. In the tissues of patients with lymph node metastasis and non lymph node metastasis of esophageal squamous cell carcinoma, the positive expression rate of Transgelin is 88.9% and 37.7% respectively, P0.05, and the difference between the two groups is statistically significant, indicating that Transgelin The expression is related to lymph node metastasis of esophageal carcinoma. Conclusion: (1) this study suggests that laser capture microdissection can effectively solve the problem of tissue heterogeneity and can better obtain more purified target cells. (2) the differential protein atlas of esophageal squamous cell carcinoma and normal esophageal epithelial cells suggests the development and development of esophageal squamous cell carcinoma. (3) the results of Western blot and immunohistochemical analysis Bmi-1, PAI-1, Cofilin-1, Transgelin are consistent with the analysis and identification of proteomics technology. (4) Bmi-l, PAI-1, Cofilin-1, and Transgelin are the four proteins in the squamous cell carcinoma of the esophagus The up-regulated expression is related to the degree of differentiation, invasion and lymphatic metastasis of esophageal squamous cell carcinoma. It may play a role in the occurrence and development of squamous cell carcinoma of the esophagus, and has a guiding significance for the malignancy and prognosis of esophageal squamous cell carcinoma, which provides a valuable theoretical basis for the molecular targeting therapy of esophageal squamous cell carcinoma and the development of new anticancer drugs.
【學(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：R735.1

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相關(guān)期刊論文 前1條

1 姜琳;孫燦林;李慧;于鴻;;胰腺癌組織中Smad7、uPA及PAI-1的表達及臨床意義[J];江蘇大學(xué)學(xué)報(醫(yī)學(xué)版);2010年06期

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本文編號：2095246