【摘要】：目的:觀察NLRC5(核苷酸結(jié)合結(jié)構(gòu)域受體家族含CARD結(jié)構(gòu)域-5)、β-catenin在胃癌組織中的表達(dá)情況。分析NLRC5與胃癌臨床病理特征及預(yù)后的關(guān)系。并初步探討NLRC5與Wnt/β-catenin信號(hào)通路間的關(guān)系。方法:收集安徽醫(yī)科大學(xué)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(shù)患者石蠟標(biāo)本93例,采用免疫組織化學(xué)染色方法(IHC)檢測(cè)胃癌組織中NLRC5和β-catenin的表達(dá)情況;分析NLRC5的表達(dá)與胃癌臨床病理特征的關(guān)系;對(duì)NLRC5表達(dá)不同的胃癌患者進(jìn)行生存分析;分析NLRC5與β-catenin的表達(dá)水平的相關(guān)性;COX多因素回歸分析NLRC5的表達(dá)是否為影響患者預(yù)后的獨(dú)立危險(xiǎn)因素。采用LipofectaminTM 2000試劑介導(dǎo)瞬時(shí)轉(zhuǎn)染p EGFP-C2-NLRC5重組質(zhì)粒入胃癌細(xì)胞株MKN-45和MGC-803中,觀察NLRC5和Wnt/β-catenin信號(hào)通路關(guān)鍵蛋白的m RNA和蛋白表達(dá)水平。q RT-PCR法和Western blot法驗(yàn)證NLRC5轉(zhuǎn)染效率。采用MTT細(xì)胞增殖和細(xì)胞劃痕實(shí)驗(yàn)對(duì)NLRC5進(jìn)行功能分析,觀察體外實(shí)驗(yàn)中NLRC5表達(dá)變化對(duì)胃癌細(xì)胞的生物學(xué)行為及Wnt/β-catenin信號(hào)通路相關(guān)蛋白的影響。結(jié)果:NLRC5在93例胃癌中的表達(dá)情況:陽性表達(dá)67例(72.04%),陰性表達(dá)26例(27.96%)。NLCR5的表達(dá)情況與胃癌的TNM分期、淋巴結(jié)轉(zhuǎn)移、復(fù)發(fā)情況密切相關(guān)。對(duì)NLCR5表達(dá)不同的患者進(jìn)行Kaplan-merier生存分析,結(jié)果顯示NLCR5陰性表達(dá)的患者預(yù)后好于陽性表達(dá)患者(P0.05)。β-catenin的表達(dá)情況:陽性表達(dá)63例(67.74%),陰性表達(dá)30例(32.26%)。spearman相關(guān)分析顯示NLRC5與β-catenin的表達(dá)呈正相關(guān),相關(guān)系數(shù)為0.814。Cox多因素回歸分析顯示,NLRC5和β-catenin的陽性表達(dá)、低TNM分期、淋巴結(jié)轉(zhuǎn)移是影響胃癌患者預(yù)后的獨(dú)立危險(xiǎn)因素。MTT細(xì)胞增殖和劃痕實(shí)驗(yàn)結(jié)果顯示,過表達(dá)NLRC5能顯著促進(jìn)胃癌細(xì)胞的增殖和遷移能力,Western blot實(shí)驗(yàn)結(jié)果顯示,過表達(dá)NLRC5能明顯提高Wnt/β-catenin信號(hào)通路中關(guān)鍵蛋白β-catenin和下游關(guān)鍵靶蛋白C-myc、MMP-7的表達(dá)水平。結(jié)論:NLRC5廣泛表達(dá)于胃癌組織中,其表達(dá)與胃癌的TNM分期和淋巴結(jié)轉(zhuǎn)移密切相關(guān)。Kaplan-merier生存分析顯示:NLRC5高表達(dá)是影響胃癌預(yù)后的危險(xiǎn)因素。COX多因素回歸分析顯示:NLRC5高表達(dá)是影響胃癌預(yù)后的獨(dú)立危險(xiǎn)因素。NLRC5與β-catenin的表達(dá)呈正相關(guān),NLRC5的表達(dá)變化影響胃癌細(xì)胞的生物學(xué)行為和Wnt/β-catenin信號(hào)通路的關(guān)鍵蛋白的表達(dá)。提示NLRC5可能通過調(diào)控Wnt/β-catenin信號(hào)通路影響胃癌的浸潤和轉(zhuǎn)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(xué)位授予單位】：安徽醫(yī)科大學(xué)
【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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