胃癌组织MYH-9蛋白表达及MYH-9 siRNA下调SGC-7901细胞MYH-9表达对细胞侵袭转移的影响  

Expression of MYH-9 in the tissues of human gastric carcinoma and the effect of siRNA-mediated inhibition of MYH-9 on invasion and metastasis in gastric carcinoma SGC-7901 cells

作　　者：曹超[1] 李太原[1] 曹传林[1] 熊凌强 刘东宁[1] 

机构地区：[1]南昌大学第一附属医院胃肠外科,江西南昌330006

出　　处：《中华肿瘤防治杂志》2016年第14期944-951,共8页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金重点项目(30930038);国家自然科学基金(81302292)

摘　　要：目的非肌性肌球蛋白重链9基因（non-muscular myosin heavy chain9，MYH-9）在多种恶性肿瘤中高表达，且预后差。检测胃癌组织MYH-9的表达及利用RNA干扰（RNA interference，RNAi）技术特异性，抑制胃癌SGC-7901细胞中MYH-9的表达对细胞侵袭和转移的影响，探讨在胃癌治疗中将MYH-9作为基因治疗靶点的价值。方法选取2009-06-01-2010-01-01南昌大学第一附属医院126例胃癌及癌旁组织。利用RNA技术将MYH-9 siRNA转染到胃癌SGC-7901细胞中，蛋白质印迹法检测MYH-9蛋白的表达，RT-PCR法检测MYH-9 mRNA的表达；运用细胞迁移和侵袭实验检测MYH-9 siRNA下调胃癌SGC-7901细胞中MYH-9的表达对细胞侵袭转移的影响。结果MYH-9在胃癌组织中的表达显著高于癌旁组织，χ2=77．83，P〈0．001。胃癌MYH-9蛋白表达与肿瘤Lauren分型（χ2=5．296，P=0．020）、肿瘤分化程度（χ2=14．39，P=0．002）、肿瘤浸润深度（χ2=5．296，P=0．026）、肿瘤淋巴结转移数（χ2=11．15,P=0．003）、脉管癌栓（χ2=5．255，P=0．022）、肿瘤远处转移状态（χ2=6．683，P=0．009）和TNM分期（χ2=10．73，P=0．002）密切相关。MYH-9蛋白表达（HR=2．825，95％CI=1．104～3．706，P=0．041）、胃癌远处转移状态（HR=1．749，95％CI=0．983～3．503，P=0．035）和胃癌淋巴结转移数（HR=3．235，95％CI=1．579～5．358，P〈0．001）可作为预测患者不良预后的独立指标。与空白对照组及阴性对照组比较，特异性siRNA可以高效抑制SGC-7901细胞MYH-9基因的表达，在mRNA水平其表达抑制率分别为87．5％（t=48．22，P〈0．001）和86．9％（t=57．30，P〈0．001）；在蛋白质水平其表达抑制率分别为52．9％（t=11．01，P〈O．001）和60．1％（t=15．53，P〈O．001）。在侵袭实验中，实验组细胞数为（49．5±13．8）个，显著低于阴性对照组（112±4．34）个（t=15．53，P〈0．001）和空白对照组（116�OBJECTIVE MYH-9 has a high expression in different tumors, and has a poor survival. This study is to detect the expression of MYH-9 in the tissue of gastric carcinoma. RNA interference （RNAi） was employed to inhibit the expression of MYH-9 in SGC-7901 cells and to evaluate the effects of MYH-9 as a target for gene therapy in gastric carci- noma. METHODS MYH-9 protein expression was examined by immunohistochemistry in 126 gastric adenocarcinoma and adjacent normal tissues. SGC-7901 cells were transfected with MYH-9 siRNA. MYH-9 protein and mRNA levels were detected using Western Blotting and RT-PCR. The migration and invasion assay were used to evaluate the effect of siR- NA-mediated inhibition of MYH-9 on invasion and metastasis in gastric carcinoma SGC-7901 cells. RESULTS MYH-9 protein was overexpressed in GC compared with the adjacent normal gastric epithelium （χ2= 77.83, P〈0. 001）. Elevated MYH-9 expression was strongly correlated with Lauren classification （χ2 = 5. 296 ,P=0. 020）, tumor differentiation（χ2 = 14.39, P = 0.002）, depth of wall invasion（χ2 = 5.296, P = 0.026）, mean no. of MLNs （χ2 = 11.15, P = 0. 003 ）, vascular tumor embolus（χ2 =5. 255,P=0. 022）, distant metastasis staus（χ2 =6. 683,P=0. 009） and clinical stage（χ2 = 10.73, P= 0. 002）. Cox regression model demonstrated that MYH-9 expression level （ HR= 2. 825,95%CI=1. 104-3. 706, P= 0. 041）, distant metastasis status（HR=1. 749,95%CI=0. 983-3. 503,P=0. 035） and mean no. of MLNs（HR=3. 235, 950, CI= 1.579=5. 358, P〈0.001） were the independent prognostic factors of gastric cancer. The specific siRNA can efficiently block the expression of MYH-9 both at mRNA and protein levels. Compared with the blank control group and negative control group, the expression inhibition rates were 87.5 % （t = 48.22, P〈 0. 001） and 86.9 % （t = 57.30, P 〈0. 001）at mRNA level, respectively. The expression inhibition rates were 52.9%0 （t=11.01,P〈0. 001） and 60.1% （t= 15.53,P〈0. 001） at potein

关 键 词：胃癌 RNA干扰 MYH-9 预后 侵袭和转移 

分 类 号：R735.2[医药卫生—肿瘤]