NLRC5在胃癌中的表达及其与胃癌临床病理特征和预后的关系
发布时间：2019-04-29 07:31
【摘要】：目的:观察NLRC5(核苷酸结合结构域受体家族含CARD结构域-5)、β-catenin在胃癌组织中的表达情况。分析NLRC5与胃癌临床病理特征及预后的关系。并初步探讨NLRC5与Wnt/β-catenin信号通路间的关系。方法:收集安徽医科大学第一附属医院2011年1月至2012年10月确诊为胃癌,根治性胃癌切除术患者石蜡标本93例,采用免疫组织化学染色方法(IHC)检测胃癌组织中NLRC5和β-catenin的表达情况;分析NLRC5的表达与胃癌临床病理特征的关系;对NLRC5表达不同的胃癌患者进行生存分析;分析NLRC5与β-catenin的表达水平的相关性;COX多因素回归分析NLRC5的表达是否为影响患者预后的独立危险因素。采用LipofectaminTM 2000试剂介导瞬时转染p EGFP-C2-NLRC5重组质粒入胃癌细胞株MKN-45和MGC-803中,观察NLRC5和Wnt/β-catenin信号通路关键蛋白的m RNA和蛋白表达水平。q RT-PCR法和Western blot法验证NLRC5转染效率。采用MTT细胞增殖和细胞划痕实验对NLRC5进行功能分析,观察体外实验中NLRC5表达变化对胃癌细胞的生物学行为及Wnt/β-catenin信号通路相关蛋白的影响。结果:NLRC5在93例胃癌中的表达情况:阳性表达67例(72.04%),阴性表达26例(27.96%)。NLCR5的表达情况与胃癌的TNM分期、淋巴结转移、复发情况密切相关。对NLCR5表达不同的患者进行Kaplan-merier生存分析,结果显示NLCR5阴性表达的患者预后好于阳性表达患者(P0.05)。β-catenin的表达情况:阳性表达63例(67.74%),阴性表达30例(32.26%)。spearman相关分析显示NLRC5与β-catenin的表达呈正相关,相关系数为0.814。Cox多因素回归分析显示,NLRC5和β-catenin的阳性表达、低TNM分期、淋巴结转移是影响胃癌患者预后的独立危险因素。MTT细胞增殖和划痕实验结果显示,过表达NLRC5能显著促进胃癌细胞的增殖和迁移能力,Western blot实验结果显示,过表达NLRC5能明显提高Wnt/β-catenin信号通路中关键蛋白β-catenin和下游关键靶蛋白C-myc、MMP-7的表达水平。结论:NLRC5广泛表达于胃癌组织中,其表达与胃癌的TNM分期和淋巴结转移密切相关。Kaplan-merier生存分析显示:NLRC5高表达是影响胃癌预后的危险因素。COX多因素回归分析显示:NLRC5高表达是影响胃癌预后的独立危险因素。NLRC5与β-catenin的表达呈正相关,NLRC5的表达变化影响胃癌细胞的生物学行为和Wnt/β-catenin信号通路的关键蛋白的表达。提示NLRC5可能通过调控Wnt/β-catenin信号通路影响胃癌的浸润和转移。
[Abstract]:Aim: to observe the expression of NLRC5 (nucleotide binding domain receptor family containing CARD domain-5) and 尾-catenin in gastric cancer. To analyze the relationship between NLRC5 and clinicopathological features and prognosis of gastric cancer. The relationship between NLRC5 and Wnt/ 尾-catenin signaling pathway was also discussed. Methods: 93 patients with gastric cancer were collected from the first affiliated Hospital of Anhui Medical University from January 2011 to October 2012. Immunohistochemical staining method (IHC) was used to detect the expression of NLRC5 and 尾-catenin in gastric cancer tissues. To analyze the relationship between the expression of NLRC5 and the clinicopathological features of gastric cancer, to analyze the survival of patients with different expression of NLRC5, to analyze the correlation between the expression of NLRC5 and 尾-catenin, and to analyze the relationship between the expression of NLRC5 and 尾-catenin. COX multivariate regression analysis showed that the expression of NLRC5 was an independent risk factor for prognosis. The recombinant plasmid p-EGFP-C2-NLRC5 was transiently transfected into gastric cancer cell lines MKN-45 and MGC-803 by LipofectaminTM 2000 reagent. The expression levels of m-RNA and protein in the key proteins of NLRC5 and Wnt/ 尾-catenin signaling pathway were observed. The transfection efficiency of NLRC5 was verified by Q-RT-PCR and Western blot methods. MTT cell proliferation and cell scratch test were used to analyze the function of NLRC5. The effects of NLRC5 expression on the biological behavior of gastric cancer cells and Wnt/ 尾-catenin signaling pathway-related proteins were observed in vitro. Results: the expression of NLRC5 was positive in 67 cases (72.04%) and negative in 26 cases (27.96%) in 93 cases of gastric carcinoma. The expression of NLCR5 was closely related to TNM stage, lymph node metastasis and recurrence of gastric cancer. Kaplan-merier survival analysis of patients with different expression of NLCR5 showed that the prognosis of patients with negative expression of NLCR5 was better than that of patients with positive expression (P0.05). The expression of 尾-catenin: 63 cases (67.74%) were positive. Spearman correlation analysis showed that there was a positive correlation between the expression of NLRC5 and 尾-catenin, the correlation coefficient was 0.814.Cox multivariate regression analysis, the positive expression of NLRC5 and 尾-catenin was found, and the low TNM stage was found. Lymph node metastasis was an independent risk factor for prognosis of patients with gastric cancer. The results of MTT cell proliferation and scratching test showed that overexpression of NLRC5 could significantly promote the proliferation and migration of gastric cancer cells., Western blot assay showed that the proliferation and migration of gastric cancer cells were significantly enhanced. Overexpression of NLRC5 can significantly increase the expression level of 尾-catenin, the downstream key target protein, and MMP-7 in the Wnt/ 尾-catenin signaling pathway. Conclusion: NLRC5 is widely expressed in gastric cancer tissues. Kaplan-merier survival analysis showed that high expression of NLRC5 was a risk factor for prognosis of gastric cancer. Cox multivariate regression analysis showed that high expression of NLRC5 was associated with prognosis of gastric cancer. Kaplan-merier survival analysis showed that high expression of NLRC5 was associated with prognosis of gastric cancer. The expression of NLRC5 was positively correlated with the expression of 尾-catenin. The expression of NLRC5 affects the biological behavior of gastric cancer cells and the expression of key proteins in Wnt/ 尾-catenin signaling pathway. These results suggest that NLRC5 may influence the invasion and metastasis of gastric cancer by regulating the Wnt/ 尾-catenin signaling pathway.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.2
本文编号：2468068
[Abstract]:Aim: to observe the expression of NLRC5 (nucleotide binding domain receptor family containing CARD domain-5) and 尾-catenin in gastric cancer. To analyze the relationship between NLRC5 and clinicopathological features and prognosis of gastric cancer. The relationship between NLRC5 and Wnt/ 尾-catenin signaling pathway was also discussed. Methods: 93 patients with gastric cancer were collected from the first affiliated Hospital of Anhui Medical University from January 2011 to October 2012. Immunohistochemical staining method (IHC) was used to detect the expression of NLRC5 and 尾-catenin in gastric cancer tissues. To analyze the relationship between the expression of NLRC5 and the clinicopathological features of gastric cancer, to analyze the survival of patients with different expression of NLRC5, to analyze the correlation between the expression of NLRC5 and 尾-catenin, and to analyze the relationship between the expression of NLRC5 and 尾-catenin. COX multivariate regression analysis showed that the expression of NLRC5 was an independent risk factor for prognosis. The recombinant plasmid p-EGFP-C2-NLRC5 was transiently transfected into gastric cancer cell lines MKN-45 and MGC-803 by LipofectaminTM 2000 reagent. The expression levels of m-RNA and protein in the key proteins of NLRC5 and Wnt/ 尾-catenin signaling pathway were observed. The transfection efficiency of NLRC5 was verified by Q-RT-PCR and Western blot methods. MTT cell proliferation and cell scratch test were used to analyze the function of NLRC5. The effects of NLRC5 expression on the biological behavior of gastric cancer cells and Wnt/ 尾-catenin signaling pathway-related proteins were observed in vitro. Results: the expression of NLRC5 was positive in 67 cases (72.04%) and negative in 26 cases (27.96%) in 93 cases of gastric carcinoma. The expression of NLCR5 was closely related to TNM stage, lymph node metastasis and recurrence of gastric cancer. Kaplan-merier survival analysis of patients with different expression of NLCR5 showed that the prognosis of patients with negative expression of NLCR5 was better than that of patients with positive expression (P0.05). The expression of 尾-catenin: 63 cases (67.74%) were positive. Spearman correlation analysis showed that there was a positive correlation between the expression of NLRC5 and 尾-catenin, the correlation coefficient was 0.814.Cox multivariate regression analysis, the positive expression of NLRC5 and 尾-catenin was found, and the low TNM stage was found. Lymph node metastasis was an independent risk factor for prognosis of patients with gastric cancer. The results of MTT cell proliferation and scratching test showed that overexpression of NLRC5 could significantly promote the proliferation and migration of gastric cancer cells., Western blot assay showed that the proliferation and migration of gastric cancer cells were significantly enhanced. Overexpression of NLRC5 can significantly increase the expression level of 尾-catenin, the downstream key target protein, and MMP-7 in the Wnt/ 尾-catenin signaling pathway. Conclusion: NLRC5 is widely expressed in gastric cancer tissues. Kaplan-merier survival analysis showed that high expression of NLRC5 was a risk factor for prognosis of gastric cancer. Cox multivariate regression analysis showed that high expression of NLRC5 was associated with prognosis of gastric cancer. Kaplan-merier survival analysis showed that high expression of NLRC5 was associated with prognosis of gastric cancer. The expression of NLRC5 was positively correlated with the expression of 尾-catenin. The expression of NLRC5 affects the biological behavior of gastric cancer cells and the expression of key proteins in Wnt/ 尾-catenin signaling pathway. These results suggest that NLRC5 may influence the invasion and metastasis of gastric cancer by regulating the Wnt/ 尾-catenin signaling pathway.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.2
【参考文献】
相关期刊论文 前2条
1 Hye Won Chung;Jong-Baeck Lim;;Role of the tumor microenvironment in the pathogenesis of gastric carcinoma[J];World Journal of Gastroenterology;2014年07期
2 段纪俊;陈万青;张思维;;中国恶性肿瘤死亡率的国际比较[J];中国社会医学杂志;2009年06期
，本文编号：2468068
本文链接：https://www.wllwen.com/yixuelunwen/zlx/2468068.html
