巨噬细胞金属弹力酶对小鼠原位结肠癌生长及COX-2表达的影响  

Effects of mouse macrophage metalloelastase on growth and cyclooxygenase-2 expression of murine orthotopic colon cancer

作　　者：石海[1] 许建明[1] 胡乃中[1] 梅俏[1] 鲍峻峻[1] 

机构地区：[1]安徽医科大学第一附属医院消化内科,安徽省合肥市230022

出　　处：《世界华人消化杂志》2007年第18期2014-2020,共7页World Chinese Journal of Digestology

摘　　要：目的:探讨巨噬细胞金属弹力酶(MME)对小鼠原位结肠癌生长、微血管生成及环氧合酶-2(COX-2)表达的影响.方法:扩增编码MME基因结构域Ⅰ和Ⅱ的cDNA片段,构建真核细胞表达载体pcDNA3.1-MME并转染小鼠CT-26结肠癌细胞.建立MME转染组及对照组小鼠原位结肠癌种植模型,观察重组MME对结肠癌生长的影响,采用Western blot方法检测肿瘤组织中的COX-2表达和微血管密度(MVD).结果:4wk后,小鼠原位结肠癌的平均体积和微血管密度,空质粒转染组小鼠和未转染组显著高于MME转染组(1151.07±35.91 mm^3,1201.13±42.15 mm^3 vs 384.83±4.76 mm^3.P<0.001;21.87±0.47,22.56±0.71 vs 8.48±0.53,P<0.001).COX-2蛋白水平在MME转染组中均显著低于对照组(2.766±1.22 vs 5.77±1.08,5.84±0.95,P<0.01).结论:转染入小鼠结肠癌细胞的MME基因通过抑制新生血管的生成和COX-2表达,从而起到抑制原位结肠癌生长的作用.AIM： To determine correlations between macrophage metalloelastase （MME）, microvessel angiogenesis and cyclooxygenase-2 （COX-2） expression in the angiogenesis of colon cancer. METHODS： A cDNA fragment coding for domains Ⅰ andⅡ of MME was transfected into MME deficient murine CT-26 colon cancer cells. The enzymatic activity of recombinant MME was confirmed by cleavage of native substrate in vitro. An orthotopic implantation model was established using MME-transfected and control cells. Tumor samples were subjected to Western blot to detect expressions of MME and COX-2. Microvessel counting was used to assess angio- genesis of murine colon tumors. RESULTS： Primary tumors with volumes and microvessel formation carrying pcDNA3.1-transfected and nontransfected cells were larger than that in the group implanted with MMEtransfected （1151.07 ± 35.91 mm^3 and 1201.13 ± 42.15 mm^3 vs 384.83 ± 4.76 mm^3, P 〈 0.001; 21.87 ± 0.47 and 22.56 ± 0.71 vs 8.48 ± 0.53, P 〈 0.001, respectively）. The expression of COX-2 protein was significantly lower in the MME-transfected group than that in the controls by Western blot analysis （2.766 ± 1.22 vs 5.77 ± 1.08 and 5.84 ± 0.95, P 〈 0.01, respectively）. CONCLUSION： The MME gene when transfected into murine colon cancer cells can effectively suppress the growth of orthotopic tumors by inhibiting vascularity and COX-2 expression.

关 键 词：巨噬细胞金属弹力酶 基因克隆 结肠癌 环氧合酶-2 动物模型 

分 类 号：R735.35[医药卫生—肿瘤]