Smad7反义寡核苷酸对人胰腺癌SW1990细胞MMP-2和TIMP-2表达及细胞增殖的影响  

Effect of Smad7 antisense oligodeoxynucleotide on the expression of MMP-2, TIMP-2 and proliferation in human pancreatic cancer cells SW1990

作　　者：于鸿[1] 黄俊星[2] 王朝夫[3] 施达仁[3] 

机构地区：[1]江苏省泰州市人民医院病理科,江苏泰州225300 [2]江苏省泰州市人民医院肿瘤科,江苏泰州225300 [3]复旦大学附属肿瘤医院病理科

出　　处：《中华胰腺病杂志》2012年第1期22-25,共4页Chinese Journal of Pancreatology

基　　金：江苏省医学重点人才基金（RC2007036）

摘　　要：目的研究Smad7反义寡核苷酸（ASODN）转染后人胰腺癌SW1990细胞基质金属蛋白酶-2（MMP-2）和基质金属蛋白酶组织抑制因子-2（TIMP-2）表达及细胞增殖的变化，探讨Smad7在胰腺癌发生、发展中的作用机制。方法经脂质体介导将Smad7ASODN转染人胰腺癌细胞株SW1990。以无义寡核苷酸（NSODN）转染、单加ASODN、单加脂质体作为对照。荧光显微镜及流式细胞仪检测转染效率；RT-PCR、蛋白质印迹法检测转染细胞Smad7、MMP-2和TIMP-2表达；噻唑蓝（MTF）比色法检测转染细胞的增殖。结果Smad7ASODN转染SW1990细胞的转染效率为81．2％，转染24h后，转染组、ASODN组和脂质体组细胞Smad7mRNA表达量分别为0．34±0．06、0．95±0．07、1．03±0．11；MMP-2mRNA表达量为0．54±0．08、1．15±0．13、1．27±O．16；TIMP-2mRNA表达量为0．26±0．07、0．72±0．13、0．78±0．17；Smad7蛋白表达量为0．14±0．03、0．294-0．05、0．284-0．07；MMP-2蛋白表达量为0．17±0．02、0．29±0．05、0．31±0．04；TIMP-2蛋白表达量为0．20±0．03、0．41±0．11、0．43±0．09。转染组均较其他各组的表达显著减少（P值均〈0．01）。转染组、ASODN组、脂质体组细胞的增殖活性分别为0．83±0．03、1．02±0．02、0．99±0．02，转染组细胞的增殖活性较其他各组显著被抑制（P值均〈0．01）。结论Smad7ASODN转染可有效抑制SW1990细胞Smad7基因的表达，并抑制MMP-2、TIMP-2表达及细胞增殖活性。Objective To investigate the effect of Smad7 antisense oligodeoxynucleotide （ASODN） on proliferation in human pancreatic cancer cell line SW1990, with a focus on the expression of matrix metalloproteinase-2 （MMP-2） and tissue inhibitor of metalloproteinase-2 （TIMP-2）. To explore the underlying mechanism of the role of Smad7 in the pathogenesis and development of pancreatic cancer. Methods Smad7 ASODN was transfected into SW1990 cells through lipofectamine. Nosense oligodeoxynucleotide （NSODN）, ASODN and lipofectamine was used as control. The transfection efficiency was assessed by fluorescence microscopy and flow cytometry. The expressions of Smad7, MMP-2 and TIMP-2 in transfected cells were detected by RT-PCR and Western blot. Cell viability was assessed by dimethyl thiazoldiphenyhetrazoliumbromide （MTT） method. Results Smad7 was expressed in SW1990 cells. The transfection efficiency of SW1990 was 81.2%. The expressions of Smad7 mRNA were 0. 34 ±0.06, 0.95± 0.07, 1.03 ± 0.11 in transfected group, ASODN and lipofectamine group ; and the expressions of MMP-2 mRNAwere 0. 54 ± 0.08, 1.15 ± 0.13, 1.27 ± 0.16 ; and the expressions of TIMP - 2 mRNA were 0. 26 ＋0.07, 0.72± 0.13, 0.78 ± 0.17, the mRNA expressions were significantly reduced in Smad7 ASODN transfected group, compared with other two groups （P 〈 0.01 ）. The expressions of Smad7 protein were 0.14± 0.03, 0.29± 0.05, 0.28± 0.07 in transfected group, ASODN and lipofectamine group; the expressions of MMP-2 protein were 0.17 ± 0.02, 0.29 ± 0.05, 0.31 ± 0.04, and the expressions of TIMP-2 protein were 0.20 ± 0.03, 0.41±0.11, 0.43 ± 0.09, the protein expressions were significantly reduced in Smad7 ASODN transfected group, compared with other two groups （P 〈 0.01 ）. The A490 values of proliferation were 0.83 ±0.03, 1.02 ± 0.02, 0.99±0.02 in transfected group, ASODN and lipofectamine group, the proliferation were significantly reduced in Smad7 ASODN transfected group, compared with other two groups （P 〈0.01 ）

关 键 词：胰腺肿瘤 SMAD7 反义寡核苷酸 基质金属蛋白酶-2 基质金属蛋白酶组织抑制因子-2 细胞增殖 

分 类 号：R735.9[医药卫生—肿瘤]