本文選題：胃癌 + NLRC��； 參考：《安徽醫(yī)科大學學報》2017年10期

【摘要】：目的觀察核苷酸結(jié)合結(jié)構(gòu)域受體家族含CARD結(jié)構(gòu)域-5(NLRC5)、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(guān)系。并初步探討NLRC5與Wnt/β-catenin信號通路間的關(guān)系。方法收集根治性胃癌切除術(shù)患者石蠟標本93例,免疫組化檢測胃癌組織中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(guān)系。采用胃癌細胞株MKN-45和MGC-803分別轉(zhuǎn)染p EGFP-C2-NLRC5重組質(zhì)粒,qRT-PCR法和Western blot法驗證NLRC5轉(zhuǎn)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的影響,采用Western blot法檢測Wnt/β-catenin信號通路相關(guān)蛋白的變化。結(jié)果 NLRC5在93例胃癌標本中陽性表達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(jié)轉(zhuǎn)移和復發(fā)情況密切相關(guān)。Kaplan-merier生存分析結(jié)果顯示NLCR5陰性表達的患者預后好于陽性表達患者(P0.05)。93例胃癌標本中β-catenin陽性表達63例(67.74%),陰性表達30例(32.26%)。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(jié)轉(zhuǎn)移是影響胃癌患者預后的獨立危險因素。胃癌細胞在過表達NLRC5后,MTT細胞增殖和劃痕實驗結(jié)果顯示,細胞的增殖和遷移能力顯著增強(P0.05,P0.01),Western blot實驗結(jié)果顯示,Wnt/β-catenin信號通路中關(guān)鍵蛋白β-catenin和下游關(guān)鍵靶蛋白C-myc、金屬基質(zhì)蛋白酶-7的表達水平明顯提高。結(jié)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(jié)轉(zhuǎn)移密切相關(guān)。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(guān)性,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(guān)鍵蛋白的表達。提示NLRC5可能通過調(diào)控Wnt/β-catenin信號通路影響胃癌的浸潤和轉(zhuǎn)移。
[Abstract]:Objective to investigate the expression of nucleotide binding domain receptor family (NLRC5, 尾 -catenin) in gastric carcinoma. To analyze the relationship between NLRC 5 and clinicopathological features and prognosis of gastric cancer. The relationship between NLRC5 and Wnt / 尾 -catenin signaling pathway was studied. Methods the expression of NLRC5 and 尾 -catenin was detected by immunohistochemistry in 93 paraffin specimens of patients with radical gastrectomy, and the relationship between the expression of NLRC5 and clinicopathological features of gastric cancer was analyzed. The transfection efficiency of pEGFP-C2-NLRC5 was verified by pEGFP-C2-NLRC5 recombinant plasmid pEGFP-C2-NLRC5 transfected with MKN-45 and MGC-803 by Western blot and RT-PCR respectively. The function of NLRC5 was analyzed by MTT cell proliferation and scratch assay. The effect of NLRC5 expression on the biological behavior of gastric cancer cells was observed in vitro. The changes of Wnt- 尾 -catenin signaling pathway related proteins were detected by Western blot assay. Results the positive expression of NLRC5 in 93 cases of gastric cancer was 72.04%, and the negative expression of NLRC5 was 27.96% in 26 cases, and the TNM stage of gastric cancer. The results of Kaplan-merier survival analysis showed that the prognosis of patients with negative expression of NLCR5 was better than that of patients with positive expression of P0.05. 93 cases of gastric cancer. 63 cases of positive expression of 尾 -catenin and 30 cases of negative expression of 尾 -catenin were analyzed by multivariate regression analysis. The positive expression of NLRC5 and 尾 -catenin, Low TNM staging and lymph node metastasis are independent risk factors for prognosis of gastric cancer patients. After overexpression of NLRC5 in gastric cancer cells, MTT cell proliferation and scratch test showed that, The cell proliferation and migration were significantly enhanced by P0.05P0.01P0.01Western blot. The results showed that the expression of 尾 -catenin, C-mycand metalloproteinase-7 in Wnt/ 尾 -catenin signaling pathway was significantly increased. Conclusion NLRC5 is widely expressed in gastric carcinoma. Kaplan-merier survival analysis showed that the high expression of 1% NLRC5 was a risk factor for the prognosis of gastric cancer. Cox multivariate regression analysis showed that the high expression of 1% NLRC5 was an independent risk factor for the prognosis of gastric cancer. The expression of NLRC5 was positively correlated with the expression of 尾 -catenin. The expression of NLRC5 affected 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 Wnt- 尾 -catenin signaling pathway.
【作者單位】： 安徽醫(yī)科大學基礎(chǔ)醫(yī)學院病理學教研室;安徽醫(yī)科大學第一附屬醫(yī)院普外科;
【基金】：國家自然科學基金(編號:81572305) 安徽省自然科學基金(編號:KJ2017A204)
【分類號】：R735.2
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本文編號：2015238