【摘要】：目的:觀察NLRC5(核苷酸結合結構域受體家族含CARD結構域-5)、β-catenin在胃癌組織中的表達情況。分析NLRC5與胃癌臨床病理特征及預后的關系。并初步探討NLRC5與Wnt/β-catenin信號通路間的關系。方法:收集安徽醫(yī)科大學第一附屬醫(yī)院2011年1月至2012年10月確診為胃癌,根治性胃癌切除術患者石蠟標本93例,采用免疫組織化學染色方法(IHC)檢測胃癌組織中NLRC5和β-catenin的表達情況;分析NLRC5的表達與胃癌臨床病理特征的關系;對NLRC5表達不同的胃癌患者進行生存分析;分析NLRC5與β-catenin的表達水平的相關性;COX多因素回歸分析NLRC5的表達是否為影響患者預后的獨立危險因素。采用LipofectaminTM 2000試劑介導瞬時轉染p EGFP-C2-NLRC5重組質粒入胃癌細胞株MKN-45和MGC-803中,觀察NLRC5和Wnt/β-catenin信號通路關鍵蛋白的m RNA和蛋白表達水平。q RT-PCR法和Western blot法驗證NLRC5轉染效率。采用MTT細胞增殖和細胞劃痕實驗對NLRC5進行功能分析,觀察體外實驗中NLRC5表達變化對胃癌細胞的生物學行為及Wnt/β-catenin信號通路相關蛋白的影響。結果:NLRC5在93例胃癌中的表達情況:陽性表達67例(72.04%),陰性表達26例(27.96%)。NLCR5的表達情況與胃癌的TNM分期、淋巴結轉移、復發(fā)情況密切相關。對NLCR5表達不同的患者進行Kaplan-merier生存分析,結果顯示NLCR5陰性表達的患者預后好于陽性表達患者(P0.05)。β-catenin的表達情況:陽性表達63例(67.74%),陰性表達30例(32.26%)。spearman相關分析顯示NLRC5與β-catenin的表達呈正相關,相關系數為0.814。Cox多因素回歸分析顯示,NLRC5和β-catenin的陽性表達、低TNM分期、淋巴結轉移是影響胃癌患者預后的獨立危險因素。MTT細胞增殖和劃痕實驗結果顯示,過表達NLRC5能顯著促進胃癌細胞的增殖和遷移能力,Western blot實驗結果顯示,過表達NLRC5能明顯提高Wnt/β-catenin信號通路中關鍵蛋白β-catenin和下游關鍵靶蛋白C-myc、MMP-7的表達水平。結論:NLRC5廣泛表達于胃癌組織中,其表達與胃癌的TNM分期和淋巴結轉移密切相關。Kaplan-merier生存分析顯示:NLRC5高表達是影響胃癌預后的危險因素。COX多因素回歸分析顯示:NLRC5高表達是影響胃癌預后的獨立危險因素。NLRC5與β-catenin的表達呈正相關,NLRC5的表達變化影響胃癌細胞的生物學行為和Wnt/β-catenin信號通路的關鍵蛋白的表達。提示NLRC5可能通過調控Wnt/β-catenin信號通路影響胃癌的浸潤和轉移。
[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.
【學位授予單位】：安徽醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R735.2

【參考文獻】

相關期刊論文 前2條

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 段紀俊;陳萬青;張思維;;中國惡性腫瘤死亡率的國際比較[J];中國社會醫(yī)學雜志;2009年06期

，

本文編號：2468068