核苷酸结合寡聚化结构域样受体3在结肠癌组织的表达及其对结肠癌细胞增殖和迁移的影响  

Expression of NOD-like receptor family pyrin domain-containing protein 3 in colorectal cancer and its role in cell proliferation and migration

作　　者：曾兵[1,2] 蔡灿锋 李英儒[3] 陈翔[1] 陈国星[1] 辛海洋[1] 曾军[1] 唐超明[1] 徐鋆耀[2] 

机构地区：[1]广州医科大学附属第六医院(清远市人民医院)胃肠外科,清远511518 [2]中山大学孙逸仙纪念医院肝胆胰外科,广州510120 [3]中山大学附属第六医院胃肠外科,广州510655

出　　处：《中华实验外科杂志》2017年第8期1394-1397,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金（81401996）;广东省自然科学基金（2015A030310099）

摘　　要：目的 观察核苷酸结合寡聚化结构域样受体3（NLRP3）在结肠癌组织的表达及对结肠癌细胞HCT116和SW480生物学行为的影响.方法 采用免疫组织化学法检测NLRP3在56例结肠癌组织及8例正常组织中蛋白的表达.实时定量反转录聚合酶链反应（RT-qPCR）检测NLRP3在20例结肠癌组织中mRNA的表达.采用Kaplan-Meier法进行生存分析,Log-rank法进行显著性检验,Cox回归进行多因素分析.采用Lipofectamine 2000进行p-NLRP3表达载体转染,RT-qPCR 和Western blot检测NLRP3 mRNA及蛋白的表达,噻唑蓝（MTT）和Transwell法检测其对结肠癌细胞增殖和迁移能力的影响.结果 56例肿瘤组织中NLRP3高表达组10例,低表达组46例,在正常结肠黏膜组织中呈阳性表达.NLRP3 mRNA在癌旁组织的表达为癌组织表达的2.51倍.NLRP3 表达与肿瘤浸润深度（P=0.024）、淋巴结转移（P=0.009）、肝转移（P=0.038）及TNM分期（P=0.024）相关.NLRP3低表达组总生存率显著低于高表达组（P=0.010）.NLRP3低表达为结肠癌预后不良独立风险因素（P=0.047）.p-NLRP3转染SW80细胞,72h对照组和p-NLRP组吸光度（A）值分别为0.63±0.06和0.91±0.03（P=0.010）,96h分别为1.07±0.05和1.48±0.05（P=0.003）.p-NLRP3转染HCT116细胞,72h对照组和p-NLRP组A值分别为0.80±0.05和1.20±0.08（P=0.005）,96h分别为1.02±0.08和1.64±0.05（P=0.003）.过表达NLRP3,SW480细胞对照组和p-NLRP组穿膜细胞数分别为（104±8）和（30±4）个（P=0.008）;HCT116细胞对照组和p-NLRP组穿膜细胞数分别为（123±6）和（36±3）个（P=0.022）.结论 NLRP3低表达与结肠癌进展密切相关,是其独立预后不良风险因素.Objective To investigate the expression of NOD-like receptor family pyrin domain-containing protein 3 （NLRP3） and its role in biological behaviors of colorectal cancer （CRC） cells.Methods The expression of NLRP3 was determined by immunohistochemistry in CRC samples and normal colon mucosal tissues.Kaplan-Meier method and Log-rank test were used to estimate survival rate.Cox proportional hazards model was used to calculate multivariate hazard ratios for the study variables.The real-time quantitative reverse transcriptase-polymerase chain reaction （RT-qPCR） and Western blotting were used to determine the mRNA and protein expression of NLRP3 respectively.Methyl thiazol tetrazolium （MTT） and Transwell invasion assay were used to determine the effect of NLRP3 on CRC cells.Results The NLRP3 expression was down-regulated in most of the CRC cells,with 46 cases low expression and 10 cases of high expression.Low NLRP3 expression was correlated with depth of invasion （P=0.024）,lymph node metastasis （P=0.009）,liver metastasis （P=0.038）,and TNM stages （P=0.024）.Patients with low NLRP3 expression had shorter overall survival than those with high expression （P=0.010）.Low NLRP3 expression was an independent prognostic factor for CRC （P=0.047）.NLRP3 over-expression could significant inhibit CRC cell proliferation and migration.The A values of SW480 in control group and p-NLRP group at 72 h were 0.63±0.06 and 0.91±0.03,respectively （P=0.010）,and at 96 h 1.07±0.05 and 1.48±0.05,respectively （P=0.003）.The optical density value （A） values of HCT116 in control group and p-NLRP group at 72nd h were 0.80±0.05 and 1.20±0.08,respectively （P=0.005）,and at 96 h 1.02±0.08 and 1.64±0.05,respectively （P=0.003）.The number of SW480 cells penetrating the membrane in control group and p-NLRP group was 104±8 and 30±4,respectively （P=0.008）.The number of HCT116 cells penetrating the membrane in control group and p-NLRP group was 123±6 and 36±3,respectively （P=0.022）.Conc

关 键 词：结肠癌 核苷酸结合寡聚化结构域样受体3 增殖 迁移 

分 类 号：R735.35[医药卫生—肿瘤]