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

【摘要】：目的:檢測ADD1在胃癌組織中的表達水平,探討在胃癌組織中ADD1的表達水平與Hedgehog信號通路之間的關系,為進一步明確胃癌的發(fā)生發(fā)展機制提供新的思路。方法:收集承德醫(yī)學院附屬醫(yī)院2013年9月至2015年6月手術切除的胃癌組織及相對應的正常胃組織各60例,這些收集來的標本都已經(jīng)過術后病理確認,證實為胃腺癌,且所有案例在手術前均未有放療及化療行為。本實驗中應用了蛋白印跡(Western Blot)和免疫組化(IHC)方法來檢測胃癌組織及相對應的正常胃組織中ADD1、IHH、PTCH1、GLI1四者蛋白的表達情況,應用熒光定量PCR(q Real-time PCR,q RT-PCR)實驗方法檢測胃癌組織及相對應的正常胃組織中ADD1、IHH、PTCH1、GLI1四者m RNA的表達情況,所得到的實驗數(shù)據(jù)用以結合臨床資料來進行統(tǒng)計學分析,以P0.05為差異有統(tǒng)計學意義。結果:1.蛋白印跡(Western Blot)實驗結果1.1ADD1、IHH、PTCH1、GLI1蛋白在胃癌組織和正常胃組織中的表達情況ADD1、IHH、PTCH1、GLI1蛋白在胃癌組織中的相對表達量均高于相對應的正常胃組織(P0.05)。1.2胃癌組織中ADD1、IHH、PTCH1、GLI1蛋白的表達情況與各項臨床病理特征之間的關系胃癌組織中ADD1、IHH、PTCH1、GLI1四者蛋白的表達均與患者的年齡、性別等因素無明顯相關關系(P0.05)。ADD1、IHH、PTCH1、GLI1蛋白在低分化的胃癌組織中的表達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蛋白的相對表達量高于浸潤深度未達到漿膜的胃癌組織(P0.05)。伴有淋巴結轉(zhuǎn)移的胃癌組織中ADD1、IHH、PTCH1、GLI1蛋白的相對表達量高于淋巴結未轉(zhuǎn)移的胃癌組織(P0.05)。2.免疫組化(IHC)實驗結果2.1 ADD1、IHH、PTCH1、GLI1蛋白在胃癌組織及正常胃組織中的表達情況ADD1、IHH、PTCH1、GLI1蛋白在胃癌組織中的表達量明顯高于正常胃組織(P0.05)。2.2胃癌組織中ADD1、IHH、PTCH1、GLI1蛋白表達情況與各項臨床病理特征之間的關系胃癌組織中ADD1、IHH、PTCH1、GLI1蛋白的表達情況均與患者的年齡、性別等因素無明顯相關關系(P0.05)。ADD1、IHH、PTCH1、GLI1蛋白的陽性表達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蛋白的陽性表達率高于浸潤深度未達到漿膜的胃癌組織(P0.05)。伴有淋巴結轉(zhuǎn)移的胃癌組織中ADD1、IHH、PTCH1、GLI1蛋白的陽性表達率高于淋巴結未轉(zhuǎn)移的胃癌組織(P0.05)。3.實時熒光定量(QReal-Time PCR)實驗結果3.1 ADD1、IHH、PTCH1、GLI1m RNA在胃癌組織及正常胃組織中的表達情況胃癌組織中ADD1、IHH、PTCH1、GLI1m RNA的表達水平均高于正常胃組織(P0.05)。3.2胃癌組織中ADD1、IHH、PTCH1、GLI1mRNA的表達水平與各項臨床病理特征之間的關系胃癌組織中ADD1、IHH、PTCH1、GLI1m RNA的表達情況均與患者的年齡、性別等因素無明顯相關關系(P0.05)。低分化的胃癌組織中ADD1、IHH、PTCH1、GLI1m RNA的表達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的表達水平高于浸潤深度未達到漿膜的胃癌組織(P0.05)。伴有淋巴結轉(zhuǎn)移的胃癌組織中ADD1、IHH、PTCH1、GLI1m RNA的表達水平高于淋巴結未轉(zhuǎn)移的胃癌組織(P0.05)。4.胃癌組織中ADD1、IHH、PTCH1、GLI1的相關關系Western Blotting實驗中ADD1蛋白的相對表達量與IHH蛋白的相對表達量呈正相關關系(r=0.611,P0.05);ADD1蛋白的相對表達量與PTCH1蛋白的相對表達量呈正相關關系(r=0.681,P0.05);ADD1蛋白的相對表達量與GLI1蛋白的相對表達量呈正相關關系(r=0.759,P0.05)。免疫組織化學實驗中ADD1蛋白的相對表達量與IHH蛋白的相對表達量呈正相關關系(r=0.532,P0.05);ADD1蛋白的相對表達量與PTCH1蛋白的相對表達量呈正相關關系(r=0.731,P0.05);ADD1蛋白的相對表達量與GLI1蛋白的相對表達量呈正相關關系(r=0.852,P0.05)。QReal Time-PCR實驗中ADD1m RNA的相對表達量與IHHm RNA的相對表達量呈正相關關系(r=0.675,P0.05);ADD1m RNA的相對表達量與PTCH1m RNA的相對表達量呈正相關關系(r=0.811,P0.05);ADD1m RNA的相對表達量與GLI1m RNA的相對表達量呈正相關關系(r=0.719,P0.05)。結論:1.ADD1在胃癌組織中的表達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)展有著密切的關系。2.ADD1的表達水平在侵及漿膜和有淋巴結轉(zhuǎn)移的胃癌組織中高于未侵及漿膜和無淋巴結轉(zhuǎn)移的胃癌組織,提示ADD1的高表達與胃癌細胞的浸潤和轉(zhuǎn)移有關。3.ADD1的表達水平與Hedgehog信號通路組件IHH、PTCH1、GLI1的表達水平呈正相關關系,提示ADD1與Hedgehog信號通路在胃癌的發(fā)生發(fā)展過程中有著密切關系。
[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.
【學位授予單位】：承德醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R735.2

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