基质金属蛋白酶26能有效促进人肺腺癌a549细胞-第三军医大学学报.doc

基质金属蛋白酶26能促进人低转移肺癌95-C细胞浸润能力 黎联1,2 孙贵银2,3 向德兵2,3 李敏2,4 黎晓强1,2 曹国强5（1、重庆市江津区中心医院呼吸内科 重庆市江津区呼吸疾病研究所、重庆市江津区中心医院肿瘤科、重庆市江津区中心医院病理科探讨MMP-26对95-C细胞浸润能力的影响及机制。构建含MMP-26 cDNA的表达质粒PUC57-mmp26，转染到MMP-26低表达的95-C细胞，并筛选稳定表达的含MMP-26 cDNA的细胞，RT-PCR检测MMP-26 mRNA、MMP-9 mRNA表达，明胶酶谱分析检测过表达MMP-26对95-C分泌MMP-9的影响，侵袭试验检测细胞体外侵袭力，双荧光免疫细胞化学染色检测MMP-26、MMP-9蛋白在细胞内定位。转染含MMP-26 cDNA表达质粒的95-C细胞MMP-26mRNA、MMP-9mRNA表达上调（p0.05）。明胶酶谱分析显示过表达MMP-26的95-C分泌MMP-9能力明显增强（p0.05），体外侵袭力增强（p0.05）。双荧光免疫细胞化学染色发现MMP-26、MMP-9在细胞内共定位。MMP-26可以促进肺癌95-C细胞的侵袭，可能部分通过调节MMP-9表达促进肺癌细胞的侵袭。 基质金属蛋白酶 95-C细胞 细胞浸润Matrix metalloproteinase-26?promotes the?low metastatic?lung cancer?95-C?cell invasion LI Lian1,2 SUN Gui-yin2,3 XIANG Debing2,3 LI Min2,4 LI Xiaoqiang1,2 CAO Guoqiang5 (1．Department of Respiratory MedicineJiangjin Institute of Respiratory Diseases, Jiangjin Centre Hospital ，Chongqing Jiangjin 402260， China . . Department of oncology，Jiangjin Centre Hospital .．Department of Pathology，Jiangjin Centre Hospital . Department of Respiratory Medicine，Chongqing 402260， China .Abstract] Objective To investigate the effects of MMP-26 on cell invasiveness in 95-C and its mechanism. Methods A full-length cDNA encoding human MMP-26 was cloned into the eukaryotic expression vector PUC57, This vector was designated PUC57-mmp26 transfected into 95-C. RT-PCR were used to detect expression of MMP-26 mRNA and MMP-9 mRNA. The secretion of MMP-9 was detected by gelatin zymography In vitro invasion assay and localization of MMP-26 and MMP-9 proteins were determined .Results Transfection of MMP-26 expressing plasmid in 95-C led to over-expression of MMP-26 mRNA and MMP-9 mRNA（P0.05）,and cell invasiveness in these cells were significantly promoted（P0.05）..The results of gelatin zymogram reveal overexpression MMP-26 increase the secretion of MMP-9（P0.05）. Double immunofluorescence staining revealed that MMP-26 and MMP-9 co-localized in MMP-26-transfected 95-C cells .Conclusion MMP-26 is involved in invasion-promoting regulating of lu