BML-111抑制Hep3B细胞迁移及机制初探  

BML-111 inhibits Hep3B cell migration via 5-LOX pathway and its mechanisms

作　　者：徐芬[1] 郝华[2] 吴德强[3] 汪庆余[2] 李里香[2] 徐静 林兰 冯雨 XU Fen;HAO Hua;WU De-qiang;WANG Qing-yu;LI Li-xiang;XU Jing;LIN Lan;FENG Yu(Dept of General Medicine,Second Affiliated Hospital of Nanchang University,Nanchang 330006,China;Dept of Pathology,Second Affiliated Hospital of Nanchang University,Nanchang 330006,China;Dept of Pharmacy,Second Affiliated Hospital of Nanchang University,Nanchang 330006,China)

机构地区：[1]南昌大学第二附属医院全科医学科,江西南昌330006 [2]南昌大学第二附属医院病理科,江西南昌330006 [3]南昌大学第二附属医院药剂科,江西南昌330006

出　　处：《中国药理学通报》2020年第6期774-778,共5页Chinese Pharmacological Bulletin

基　　金：江西省自然科学基金资助项目(No 20171BAB205054)。

摘　　要：目的研究BML-111对Hep3B细胞迁移的作用并对其机制进行初步探讨。方法分别用BML-111和(或)Boc-2(BML-111受体阻断剂)刺激Hep3B细胞,观察两者对细胞形态的影响;Transwell实验研究BML-111和(或)Boc-2对Hep3B细胞迁移的影响;免疫荧光及Western blot检测BML-111和(或)Boc-2对Hep3B细胞5-脂加氧酶(5-lipoxygenase,5-LOX)核转位的影响,RT-PCR检测BML-111和(或)Boc-2对Hep3B细胞MMP-2及MMP-9基因表达的影响。结果空白组Hep3B细胞伸出许多突起,排列不紧密,即肝癌细胞发生了上皮-间质转化(epithelial-mesenchymal transition,EMT),而加入BML-111后,细胞恢复了上皮细胞的形态特点,Boc-2能阻断其作用;Transwell实验显示,BML-111能明显抑制Hep3B细胞迁移,而Boc-2能阻断其作用。免疫荧光显示,BML-111能明显降低5-LOX蛋白的表达,并促进其向胞质转位,而Boc-2能增加5-LOX蛋白的表达,并促进其向核转位;RT-PCR实验发现,BML-111能下调MMP-2及9 mRNA的表达,而Boc-2能逆转这种效应。结论BML-111可能通过抑制5-LOX进而抑制Hep3B细胞迁移。Aim To investigate the effect of BML-111 on the migration of Hep3B cell lines and possible un-derlying mechanisms.Methods Cells were divided into three groups:Control,BML-111 and BML-111+Boc-2(the specific inhibitor of FPR2)groups.Cell morphology was observed after cells were treated with different conditions.The effect of BML-111 on migration of Hep3B cells was detected by transwell assay.5-LOX protein expression and location were detected by western blot and immunofluorescence,and in RT-PCR experiments,MMP-2 and MMP-9 mRNA expressions were detected.Results For control groups,cells showed loss of epithelial morphology,became dissociated from the epithelial clusters and acquired a mesenchymal phenotypea process called the epithelial-mesenchymal transition(EMT);however,Boc-2 could block the effect of BML-111,and cells reverted to morphology of control.However,Boc-2 could block the effect of BML-111.BML-111 effectively inhibited the migration of Hep3B cells in transwell assay.BML-111 decreased 5-LOX expression and promoted it to translocate to cytoplasm,while Boc-2 could increase its expression and promote it to translocate to nucleus.Moreover,BML-111 could decrease 5-LOX protein expression and its downstream molecule,such as MMPs.Conclusion BML-111 effectively inhibits Hep3B cell migration via 5-LOX pathway.

关 键 词：5-脂加氧酶 BML-111 迁移 上皮-间质转化 转位 基质金属蛋白酶 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R345[医药卫生—基础医学]