可溶性活化matriptase-2抑制乳腺癌细胞侵袭  

Soluble activated matriptase-2 inhibits invasion of breast cancer cells

作　　者：杨剑峰[1] 左斌[1] 陈颖[1] 何杨[1] 

机构地区：[1]苏州大学教育部血液与血管疾病诊疗药物技术工程中心,江苏省215006

出　　处：《江苏医药》2016年第23期2540-2542,F0002,共4页Jiangsu Medical Journal

摘　　要：目的探讨可溶性活化的matriptase-2对乳腺癌细胞的侵袭作用及其可能机制。方法构建表达matriptase-2胞外区片段且与C末端V5标签形成融合表达的真核表达质粒,转染CHO卵巢细胞,潮霉素筛选稳定表达重组蛋白的细胞克隆,收集上清液过镍琼脂糖凝胶柱。采用Western blot法鉴定matriptase-2活化,Transwell侵袭实验观察重组可溶性活化的matriptase-2对MDA-MB-231乳腺癌细胞侵袭能力的影响,Western blot法检测MMP-9的表达。结果建立了稳定分泌matritpase-2蛋白的CHO细胞克隆,可溶性matritpase-2蛋白过镍柱后自身活化。可溶性活化的matripase-2蛋白能显著抑制MDA-MB-231细胞的侵袭能力以及MMP-9的表达。结论可溶性活化的matriptase-2可能通过降低MMP-9的表达,从而抑制乳腺癌细胞的体外侵袭作用。Objective To investigate the effect and underlying mechanism of soluble activated matriptase-2 on invasion of breast cancer cells .Methods Eukaryotic expression plasmid coding extracellular region of matriptase-2 and C-terminal V5 tag fusion gene was constructed and then transfected into CHO ovary cells for selecting stable expression by hygromycin .Cell culture supernatant was collected and treated by Ni2＋ sepharose beads ,which followed by Western blot for identification of auto-activation .The effect of soluble activated matriptase-2 on invasion of MDA-MB-231 breast cancer cells was determined by Transwell assay ,and MMP-9 expression was detected by Western blot .Results Cell clone for stable secretion of soluble matriptase-2 was successfully constructed .Soluble matriptase-2 was auto-activated after the supernatant flowed through Ni2＋sepharose column .Soluble activated matriptase-2 could significantly inhibit invasion of MDA-MB-231 cells and MMP-9 expression .Conclusion Soluble activated matriptase-2 inhibits invasion of breast cancer cells probably via down-regulating MMP-9 expression .

关 键 词：Matriptase-2 乳腺癌 侵袭 

分 类 号：R737[医药卫生—肿瘤]