本文選題：胃癌 + ADD1��； 參考：《承德醫(yī)學(xué)院》2017年碩士論文

【摘要】：目的:檢測ADD1在胃癌組織中的表達(dá)水平,探討在胃癌組織中ADD1的表達(dá)水平與Hedgehog信號(hào)通路之間的關(guān)系,為進(jìn)一步明確胃癌的發(fā)生發(fā)展機(jī)制提供新的思路。方法:收集承德醫(yī)學(xué)院附屬醫(yī)院2013年9月至2015年6月手術(shù)切除的胃癌組織及相對(duì)應(yīng)的正常胃組織各60例,這些收集來的標(biāo)本都已經(jīng)過術(shù)后病理確認(rèn),證實(shí)為胃腺癌,且所有案例在手術(shù)前均未有放療及化療行為。本實(shí)驗(yàn)中應(yīng)用了蛋白印跡(Western Blot)和免疫組化(IHC)方法來檢測胃癌組織及相對(duì)應(yīng)的正常胃組織中ADD1、IHH、PTCH1、GLI1四者蛋白的表達(dá)情況,應(yīng)用熒光定量PCR(q Real-time PCR,q RT-PCR)實(shí)驗(yàn)方法檢測胃癌組織及相對(duì)應(yīng)的正常胃組織中ADD1、IHH、PTCH1、GLI1四者m RNA的表達(dá)情況,所得到的實(shí)驗(yàn)數(shù)據(jù)用以結(jié)合臨床資料來進(jìn)行統(tǒng)計(jì)學(xué)分析,以P0.05為差異有統(tǒng)計(jì)學(xué)意義。結(jié)果:1.蛋白印跡(Western Blot)實(shí)驗(yàn)結(jié)果1.1ADD1、IHH、PTCH1、GLI1蛋白在胃癌組織和正常胃組織中的表達(dá)情況ADD1、IHH、PTCH1、GLI1蛋白在胃癌組織中的相對(duì)表達(dá)量均高于相對(duì)應(yīng)的正常胃組織(P0.05)。1.2胃癌組織中ADD1、IHH、PTCH1、GLI1蛋白的表達(dá)情況與各項(xiàng)臨床病理特征之間的關(guān)系胃癌組織中ADD1、IHH、PTCH1、GLI1四者蛋白的表達(dá)均與患者的年齡、性別等因素?zé)o明顯相關(guān)關(guān)系(P0.05)。ADD1、IHH、PTCH1、GLI1蛋白在低分化的胃癌組織中的表達(dá)水平均高于高中分化的胃癌組織(P0.05)。TNM分期為ⅠⅡ期的胃癌組織中ADD1、IHH、PTCH1、GLI1蛋白表達(dá)水平低于TNM分期為ⅢⅣ期的胃癌組織(P0.05)。伴有漿膜浸潤的胃癌組織中ADD1、IHH、PTCH1、GLI1蛋白的相對(duì)表達(dá)量高于浸潤深度未達(dá)到漿膜的胃癌組織(P0.05)。伴有淋巴結(jié)轉(zhuǎn)移的胃癌組織中ADD1、IHH、PTCH1、GLI1蛋白的相對(duì)表達(dá)量高于淋巴結(jié)未轉(zhuǎn)移的胃癌組織(P0.05)。2.免疫組化(IHC)實(shí)驗(yàn)結(jié)果2.1 ADD1、IHH、PTCH1、GLI1蛋白在胃癌組織及正常胃組織中的表達(dá)情況ADD1、IHH、PTCH1、GLI1蛋白在胃癌組織中的表達(dá)量明顯高于正常胃組織(P0.05)。2.2胃癌組織中ADD1、IHH、PTCH1、GLI1蛋白表達(dá)情況與各項(xiàng)臨床病理特征之間的關(guān)系胃癌組織中ADD1、IHH、PTCH1、GLI1蛋白的表達(dá)情況均與患者的年齡、性別等因素?zé)o明顯相關(guān)關(guān)系(P0.05)。ADD1、IHH、PTCH1、GLI1蛋白的陽性表達(dá)率在低分化的胃癌組織中高于高中分化的胃癌組織(P0.05)。TNM分期為ⅠⅡ期的胃癌組織中ADD1、IHH、PTCH1、GLI1蛋白的陽性表達(dá)率低于TNM分期為ⅢⅣ期的胃癌組織(P0.05)。伴有漿膜浸潤的胃癌組織中ADD1、IHH、PTCH1、GLI1蛋白的陽性表達(dá)率高于浸潤深度未達(dá)到漿膜的胃癌組織(P0.05)。伴有淋巴結(jié)轉(zhuǎn)移的胃癌組織中ADD1、IHH、PTCH1、GLI1蛋白的陽性表達(dá)率高于淋巴結(jié)未轉(zhuǎn)移的胃癌組織(P0.05)。3.實(shí)時(shí)熒光定量(QReal-Time PCR)實(shí)驗(yàn)結(jié)果3.1 ADD1、IHH、PTCH1、GLI1m RNA在胃癌組織及正常胃組織中的表達(dá)情況胃癌組織中ADD1、IHH、PTCH1、GLI1m RNA的表達(dá)水平均高于正常胃組織(P0.05)。3.2胃癌組織中ADD1、IHH、PTCH1、GLI1mRNA的表達(dá)水平與各項(xiàng)臨床病理特征之間的關(guān)系胃癌組織中ADD1、IHH、PTCH1、GLI1m RNA的表達(dá)情況均與患者的年齡、性別等因素?zé)o明顯相關(guān)關(guān)系(P0.05)。低分化的胃癌組織中ADD1、IHH、PTCH1、GLI1m RNA的表達(dá)水平高于高中分化的胃癌組織(P0.05)。TNM分期為ⅠⅡ期的胃癌組織中ADD1、IHH、PTCH1、GLI1m RNA的表達(dá)水平低于TNM分期為ⅢⅣ期的胃癌組織(P0.05)。伴有漿膜浸潤的胃癌組織中ADD1、IHH、PTCH1、GLI1m RNA的表達(dá)水平高于浸潤深度未達(dá)到漿膜的胃癌組織(P0.05)。伴有淋巴結(jié)轉(zhuǎn)移的胃癌組織中ADD1、IHH、PTCH1、GLI1m RNA的表達(dá)水平高于淋巴結(jié)未轉(zhuǎn)移的胃癌組織(P0.05)。4.胃癌組織中ADD1、IHH、PTCH1、GLI1的相關(guān)關(guān)系Western Blotting實(shí)驗(yàn)中ADD1蛋白的相對(duì)表達(dá)量與IHH蛋白的相對(duì)表達(dá)量呈正相關(guān)關(guān)系(r=0.611,P0.05);ADD1蛋白的相對(duì)表達(dá)量與PTCH1蛋白的相對(duì)表達(dá)量呈正相關(guān)關(guān)系(r=0.681,P0.05);ADD1蛋白的相對(duì)表達(dá)量與GLI1蛋白的相對(duì)表達(dá)量呈正相關(guān)關(guān)系(r=0.759,P0.05)。免疫組織化學(xué)實(shí)驗(yàn)中ADD1蛋白的相對(duì)表達(dá)量與IHH蛋白的相對(duì)表達(dá)量呈正相關(guān)關(guān)系(r=0.532,P0.05);ADD1蛋白的相對(duì)表達(dá)量與PTCH1蛋白的相對(duì)表達(dá)量呈正相關(guān)關(guān)系(r=0.731,P0.05);ADD1蛋白的相對(duì)表達(dá)量與GLI1蛋白的相對(duì)表達(dá)量呈正相關(guān)關(guān)系(r=0.852,P0.05)。QReal Time-PCR實(shí)驗(yàn)中ADD1m RNA的相對(duì)表達(dá)量與IHHm RNA的相對(duì)表達(dá)量呈正相關(guān)關(guān)系(r=0.675,P0.05);ADD1m RNA的相對(duì)表達(dá)量與PTCH1m RNA的相對(duì)表達(dá)量呈正相關(guān)關(guān)系(r=0.811,P0.05);ADD1m RNA的相對(duì)表達(dá)量與GLI1m RNA的相對(duì)表達(dá)量呈正相關(guān)關(guān)系(r=0.719,P0.05)。結(jié)論:1.ADD1在胃癌組織中的表達(dá)水平高于正常胃組織,在低分化的胃癌組織中高于高中分化的胃癌組織,提示ADD1與胃癌的發(fā)生發(fā)展有著密切的關(guān)系。2.ADD1的表達(dá)水平在侵及漿膜和有淋巴結(jié)轉(zhuǎn)移的胃癌組織中高于未侵及漿膜和無淋巴結(jié)轉(zhuǎn)移的胃癌組織,提示ADD1的高表達(dá)與胃癌細(xì)胞的浸潤和轉(zhuǎn)移有關(guān)。3.ADD1的表達(dá)水平與Hedgehog信號(hào)通路組件IHH、PTCH1、GLI1的表達(dá)水平呈正相關(guān)關(guān)系,提示ADD1與Hedgehog信號(hào)通路在胃癌的發(fā)生發(fā)展過程中有著密切關(guān)系。
[Abstract]:Objective: to detect the expression level of ADD1 in gastric cancer tissue and to explore the relationship between the expression level of ADD1 and the Hedgehog signaling pathway in gastric cancer tissue, and to provide a new way of thinking for further clarifying the pathogenesis and development of gastric cancer. Methods: to collect the gastric cancer tissues and relative in the Affiliated Hospital of Chengde Medical College from September 2013 to June 2015. There were 60 cases of normal gastric tissue. These collected specimens have been confirmed by pathology, confirmed as gastric adenocarcinoma, and all cases were not treated with radiotherapy and chemotherapy before operation. In this experiment, Western Blot and immunohistochemical (IHC) methods were used to detect ADD1, IHH, P in gastric cancer tissue and corresponding normal gastric tissues. The expression of TCH1, GLI1 four protein and fluorescence quantitative PCR (Q Real-time PCR, Q RT-PCR) were used to detect the expression of ADD1, IHH, PTCH1, GLI1 four in normal gastric tissue, and the experimental data were used for statistical analysis in combination with clinical data. Results: 1. protein blot (Western Blot) experimental results of 1.1ADD1, IHH, PTCH1, GLI1 protein expression in gastric cancer tissue and normal gastric tissue, ADD1, IHH, PTCH1, GLI1 protein in gastric cancer tissues are higher than the corresponding normal gastric tissue (P0.05).1.2 gastric cancer tissues The expression of ADD1, IHH, PTCH1, GLI1 four protein in gastric carcinoma tissue was not associated with the age and sex of the patients (P0.05).ADD1, IHH, PTCH1, and GLI1 protein expression levels in the low differentiated gastric carcinoma tissues were higher than those of high school differentiated gastric cancer tissue (P0.05).TNM stage for stage I and II. The expression level of ADD1, IHH, PTCH1, GLI1 protein in cancer tissues is lower than that of TNM stage of stage III IV gastric carcinoma (P0.05). The relative expression of ADD1, IHH, PTCH1, GLI1 protein in the gastric carcinoma with serous infiltration is higher than that in the gastric carcinoma without the infiltration depth (P0.05). The expression of 2.1 ADD1, IHH, PTCH1, GLI1 protein in gastric carcinoma tissue and normal gastric tissue was higher than that of non metastatic lymph node (P0.05).2. immunohistochemistry (IHC). IHH, PTCH1, GLI1 protein expression in gastric carcinoma tissues was significantly higher than that of normal gastric tissue (P0.05). The relationship between the expression of GLI1 protein and various clinicopathological features was related to the expression of ADD1, IHH, PTCH1, and GLI1 protein in gastric cancer tissues. There was no significant correlation between the age and sex factors of the patients (P0.05).ADD1, IHH, PTCH1, GLI1 protein positive rate in the low differentiated gastric cancer tissues (P0.05). The positive expression rate of ADD1, IHH, PTCH1, GLI1 protein in TNM stage of stage I and II gastric carcinoma was lower than that of TNM staging stage III IV gastric carcinoma (P0.05). The positive rate of ADD1, IHH, PTCH1, GLI1 protein in the gastric carcinoma with serous infiltration was higher than that of the gastric cancer tissue (P0.05) without the depth of infiltration. The positive expression rate of ADD1, IHH, PTCH1, GLI1 protein was higher than that of non metastatic lymph nodes (P0.05).3. real-time quantitative fluorescence quantitative (QReal-Time PCR) experimental results of 3.1 ADD1, IHH, PTCH1. The expression level of ADD1, IHH, PTCH1, GLI1mRNA in tissue (P0.05).3.2 gastric carcinoma and its clinicopathological features are related to the expression of ADD1, IHH, PTCH1, GLI1m RNA in gastric cancer tissue. There is no significant correlation between the age and sex factors of the patients. The expression level of ADD1, IHH, PTCH1, GLI1m RNA in gastric cancer tissues of stage I and II stage of gastric cancer tissue (P0.05) is lower than that in stage I and II stage of gastric cancer tissue (P0.05). The level of ADD1, IHH, PTCH1, and PTCH1, is higher than that of the serous infiltrating gastric carcinoma. The expression level of ADD1, IHH, PTCH1, GLI1m RNA in gastric carcinoma with lymph node metastasis is higher than that of ADD1, IHH, PTCH1, GLI1 in gastric cancer tissue (P0.05) without metastasis of lymph nodes. The relative expression of protein was positively correlated with the relative expression of PTCH1 protein (r=0.681, P0.05); the relative expression of ADD1 protein was positively correlated with the relative expression of GLI1 protein (r=0.759, P0.05). The relative expression of ADD1 protein in the immunohistochemical experiment was positively correlated with the relative expression of IHH protein (r=0.532, P0.0) 5) the relative expression of ADD1 protein was positively correlated with the relative expression of PTCH1 protein (r=0.731, P0.05), and the relative expression of ADD1 protein was positively correlated with the relative expression of GLI1 protein (r=0.852, P0.05).QReal Time-PCR experiment. 05) the relative expression of ADD1m RNA was positively correlated with the relative expression of PTCH1m RNA (r=0.811, P0.05); the relative expression of ADD1m RNA was positively correlated with the relative expression of GLI1m RNA (r=0.719, P0.05). Conclusion: the expression level in gastric cancer tissues is higher than that of normal gastric tissue, and it is higher than high school in low differentiated gastric cancer tissue. The differentiated gastric carcinoma suggests that ADD1 has a close relationship with the development of gastric cancer. The expression level of.2.ADD1 is higher than that of the serosa and lymph node metastases, which are higher than that of the non invasive and non lymph node metastasis gastric cancer tissues, suggesting that the high expression of ADD1 is related to the level of expression of.3.ADD1 and the level of He in the invasion and metastasis of gastric cancer cells and He The expression level of the dgehog signaling pathway components IHH, PTCH1, and GLI1 is positively correlated, suggesting that the ADD1 and Hedgehog signaling pathways are closely related to the development of gastric cancer.
【學(xué)位授予單位】：承德醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.2

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