弗林蛋白酶抑制剂对乳腺癌MCF-7细胞生长和迁移的影响

目的 探讨乳腺癌转移的机制,为深入研究乳腺癌发生、发展机制提供理论基础.方法 应用不同浓度的弗林蛋白酶（Furin）抑制剂α1-PDX处理乳腺癌MCF-7细胞.用四甲基偶氮唑蓝（MTT）和克隆形成实验检测Furin抑制剂对MCF-7细胞增殖和克隆形成的影响.单层细胞迁移实验和Transwell实验检测MCF-7细胞迁移和浸润能力.Hoechst 33342/PI双染法检测细胞凋亡.酶联免疫吸附法检测细胞培养液中基质金属蛋白酶2（MMP-2）和MMP-9蛋白水平.Western blot检测细胞迁移相关蛋白MT1-MMP、血管内皮生长因子（VEGF）-C和VEGF-D水平.结果 不同浓度α1-PDX作用MCF-7细胞48 h以上时,细胞的生长受到抑制,集落形成降低,细胞凋亡率升高.但在低浓度情况下对细胞迁移和侵袭起抑制作用.α1-PDX降低了细胞内MT1-MMP、VEGF-C、VEGF-D的表达,同时细胞培养液上清中MMP-2和MMP-9浓度也低于对照组.结论 Furin抑制剂通过抑制乳腺癌MCF-7细胞MMP及VEGF表达抑制肿瘤的迁移能力.

Effect of Furin proteinase inhibitor on growth and metastasis of breast cancer cells MCF-7

Objective To investigate the mechanisms of breast cancer metastasis and to establish basis for the further study of breast cancer.Methods MCF-7 cells were treated with different concentrations of Furin inhibitor α1-PDX for dose dependent and time course analysis.The MCF-7 cell proliferation and clone formation were assayed with MTT and colony.Wound Healing and Transwell assay were used to detect MCF-7 cell migration and invasion ability.Hoechst 33342/PI double staining was used to detect cell apoptosis.The enzyme-linked immunosorbent assay was used to detect the MMP-2 and MMP-9 levels in cell culture medium.Cell migration associated proteins （MT1-MMP,VEGF-C and VEGF-D） were monitored with Western blot.Results Cell growth and colony formation ability were inhibited by the Furin inhibitor treatment for 48 h,that accordance with cell apoptosis increasing.However,cell migration and invasion were inhibited significantly in the case of a low concentration.The expression of MT1-MMP,VEGF-C and VEGF-D decreased significantly compared to control group,which associated with MMP-2 and MMP-9 concentrations.Conclusion The migration of MCF-7 cells reduce upon Furin inhibitors treatment through down-regulation the expression of MMP and VEGF expression.