ADD1在胃癌组织中的表达及与Hedgehog信号通路关系的研究

发布时间：2018-05-28 16:35

本文选题：胃癌 + ADD1　；参考：《承德医学院》2017年硕士论文

【摘要】：目的:检测ADD1在胃癌组织中的表达水平,探讨在胃癌组织中ADD1的表达水平与Hedgehog信号通路之间的关系,为进一步明确胃癌的发生发展机制提供新的思路。方法:收集承德医学院附属医院2013年9月至2015年6月手术切除的胃癌组织及相对应的正常胃组织各60例,这些收集来的标本都已经过术后病理确认,证实为胃腺癌,且所有案例在手术前均未有放疗及化疗行为。本实验中应用了蛋白印迹(Western Blot)和免疫组化(IHC)方法来检测胃癌组织及相对应的正常胃组织中ADD1、IHH、PTCH1、GLI1四者蛋白的表达情况,应用荧光定量PCR(q Real-time PCR,q RT-PCR)实验方法检测胃癌组织及相对应的正常胃组织中ADD1、IHH、PTCH1、GLI1四者m RNA的表达情况,所得到的实验数据用以结合临床资料来进行统计学分析,以P0.05为差异有统计学意义。结果:1.蛋白印迹(Western Blot)实验结果1.1ADD1、IHH、PTCH1、GLI1蛋白在胃癌组织和正常胃组织中的表达情况ADD1、IHH、PTCH1、GLI1蛋白在胃癌组织中的相对表达量均高于相对应的正常胃组织(P0.05)。1.2胃癌组织中ADD1、IHH、PTCH1、GLI1蛋白的表达情况与各项临床病理特征之间的关系胃癌组织中ADD1、IHH、PTCH1、GLI1四者蛋白的表达均与患者的年龄、性别等因素无明显相关关系(P0.05)。ADD1、IHH、PTCH1、GLI1蛋白在低分化的胃癌组织中的表达水平均高于高中分化的胃癌组织(P0.05)。TNM分期为ⅠⅡ期的胃癌组织中ADD1、IHH、PTCH1、GLI1蛋白表达水平低于TNM分期为ⅢⅣ期的胃癌组织(P0.05)。伴有浆膜浸润的胃癌组织中ADD1、IHH、PTCH1、GLI1蛋白的相对表达量高于浸润深度未达到浆膜的胃癌组织(P0.05)。伴有淋巴结转移的胃癌组织中ADD1、IHH、PTCH1、GLI1蛋白的相对表达量高于淋巴结未转移的胃癌组织(P0.05)。2.免疫组化(IHC)实验结果2.1 ADD1、IHH、PTCH1、GLI1蛋白在胃癌组织及正常胃组织中的表达情况ADD1、IHH、PTCH1、GLI1蛋白在胃癌组织中的表达量明显高于正常胃组织(P0.05)。2.2胃癌组织中ADD1、IHH、PTCH1、GLI1蛋白表达情况与各项临床病理特征之间的关系胃癌组织中ADD1、IHH、PTCH1、GLI1蛋白的表达情况均与患者的年龄、性别等因素无明显相关关系(P0.05)。ADD1、IHH、PTCH1、GLI1蛋白的阳性表达率在低分化的胃癌组织中高于高中分化的胃癌组织(P0.05)。TNM分期为ⅠⅡ期的胃癌组织中ADD1、IHH、PTCH1、GLI1蛋白的阳性表达率低于TNM分期为ⅢⅣ期的胃癌组织(P0.05)。伴有浆膜浸润的胃癌组织中ADD1、IHH、PTCH1、GLI1蛋白的阳性表达率高于浸润深度未达到浆膜的胃癌组织(P0.05)。伴有淋巴结转移的胃癌组织中ADD1、IHH、PTCH1、GLI1蛋白的阳性表达率高于淋巴结未转移的胃癌组织(P0.05)。3.实时荧光定量(QReal-Time PCR)实验结果3.1 ADD1、IHH、PTCH1、GLI1m RNA在胃癌组织及正常胃组织中的表达情况胃癌组织中ADD1、IHH、PTCH1、GLI1m RNA的表达水平均高于正常胃组织(P0.05)。3.2胃癌组织中ADD1、IHH、PTCH1、GLI1mRNA的表达水平与各项临床病理特征之间的关系胃癌组织中ADD1、IHH、PTCH1、GLI1m RNA的表达情况均与患者的年龄、性别等因素无明显相关关系(P0.05)。低分化的胃癌组织中ADD1、IHH、PTCH1、GLI1m RNA的表达水平高于高中分化的胃癌组织(P0.05)。TNM分期为ⅠⅡ期的胃癌组织中ADD1、IHH、PTCH1、GLI1m RNA的表达水平低于TNM分期为ⅢⅣ期的胃癌组织(P0.05)。伴有浆膜浸润的胃癌组织中ADD1、IHH、PTCH1、GLI1m RNA的表达水平高于浸润深度未达到浆膜的胃癌组织(P0.05)。伴有淋巴结转移的胃癌组织中ADD1、IHH、PTCH1、GLI1m RNA的表达水平高于淋巴结未转移的胃癌组织(P0.05)。4.胃癌组织中ADD1、IHH、PTCH1、GLI1的相关关系Western Blotting实验中ADD1蛋白的相对表达量与IHH蛋白的相对表达量呈正相关关系(r=0.611,P0.05);ADD1蛋白的相对表达量与PTCH1蛋白的相对表达量呈正相关关系(r=0.681,P0.05);ADD1蛋白的相对表达量与GLI1蛋白的相对表达量呈正相关关系(r=0.759,P0.05)。免疫组织化学实验中ADD1蛋白的相对表达量与IHH蛋白的相对表达量呈正相关关系(r=0.532,P0.05);ADD1蛋白的相对表达量与PTCH1蛋白的相对表达量呈正相关关系(r=0.731,P0.05);ADD1蛋白的相对表达量与GLI1蛋白的相对表达量呈正相关关系(r=0.852,P0.05)。QReal Time-PCR实验中ADD1m RNA的相对表达量与IHHm RNA的相对表达量呈正相关关系(r=0.675,P0.05);ADD1m RNA的相对表达量与PTCH1m RNA的相对表达量呈正相关关系(r=0.811,P0.05);ADD1m RNA的相对表达量与GLI1m RNA的相对表达量呈正相关关系(r=0.719,P0.05)。结论:1.ADD1在胃癌组织中的表达水平高于正常胃组织,在低分化的胃癌组织中高于高中分化的胃癌组织,提示ADD1与胃癌的发生发展有着密切的关系。2.ADD1的表达水平在侵及浆膜和有淋巴结转移的胃癌组织中高于未侵及浆膜和无淋巴结转移的胃癌组织,提示ADD1的高表达与胃癌细胞的浸润和转移有关。3.ADD1的表达水平与Hedgehog信号通路组件IHH、PTCH1、GLI1的表达水平呈正相关关系,提示ADD1与Hedgehog信号通路在胃癌的发生发展过程中有着密切关系。
[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.
【学位授予单位】：承德医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.2

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