姜黄素通过下调FoxM1、NEDD4及β-catenin抑制黑素瘤细胞增殖、迁移及侵袭  

Curcumin inhibits the proliferation,migration and invasion of melanoma cells by down-regulating FoxM1,NEDD4 and β-catenin

作　　者：张泽鹏 沃晓嫚[2] 刘添 秦文欣 林雨丝 王永晨[3] ZHANG Ze-peng;WO Xiao-man;LIU Tian;QIN Wen-xin;LIN Yu-si;WANG Yongchen(Department of Clinical Affair,The Second Affiliated Hospital of Harbin Medical University;Department of Cell Biology,Harbin Medical University;Department of Dermatology,The Second Affiliated Hospital of Harbin Medical University,Harbin 150081,China)

机构地区：[1]哈尔滨医科大学附属第二医院医务部 [2]哈尔滨医科大学细胞生物学教研室 [3]哈尔滨医科大学附属第二医院皮肤科,黑龙江哈尔滨150081

出　　处：《哈尔滨医科大学学报》2022年第5期419-425,共7页Journal of Harbin Medical University

基　　金：黑龙江省自然科学基金面上项目(H2018024)

摘　　要：目的研究姜黄素对黑素瘤细胞增殖、迁移和侵袭的影响及其分子机制。方法将两种人黑素瘤细胞系(A375和A875)分别用不同浓度的姜黄素处理,应用MTT、划痕、Transwell实验观察姜黄素对黑素瘤细胞增殖、迁移和侵袭的影响,利用蛋白印迹技术测定细胞周期蛋白A2(CyclinA2)、细胞周期蛋白D1(CyclinD1)、叉头框蛋白M1(forkhead box protein M1,FoxM1)、神经前体细胞表达发育下调蛋白4(neural precursor cell expressed developmentally down-regulated protein 4,NEDD4)、β-连环蛋白(β-catenin)的表达情况。结果MTT与集落形成实验结果显示,姜黄素处理后细胞相对活力下降,细胞增殖能力显著下降,且具有剂量依赖性(P<0.001)。划痕和侵袭实验结果显示,姜黄素处理后黑素瘤细胞迁移相对距离明显增加,侵袭的相对细胞数明显减少,差异具有统计学意义(P<0.001;P<0.05)。Western blot结果显示,姜黄素处理后Cyclin D1和Cyclin A2、FoxM1表达水平降低(P<0.001;P<0.05);过表达FoxM1促进NEDD4表达,敲减FoxM1则抑制NEDD4表达(P<0.001)。与对照组比较,姜黄素处理后细胞核中β-catenin水平明显降低,而细胞质中β-catenin水平明显升高(P<0.001)。结论姜黄素可通过下调FoxM1/NEDD4和β-catenin抑制黑素瘤细胞的增殖、迁移和侵袭。Objective To study the effect of curcumin on the proliferation,migration and invasion of melanoma cells and its molecular mechanism.Methods Human melanoma cell lines(A375 and A875)were treated with curcumin at various concentrations.The effects of curcumin on the proliferation,migration and invasion of melanoma was observed by MTT,Wound-healing and Transwell assays.The protein expression levels of Cyclin A2,Cyclin D1,forkhead box protein M1(FoxM1),neural precursor cell expressed developmentally down-regulated protein 4(NEDD4)andβ-catenin were detected by Western blot.Results MTT and colony formation experiments showed that relative cell viability and cell proliferation was decreased significantly in a dose-dependent manner after curcumin treatment(P<0.001).The results of Wound-healing and invasion experiments showed that the relative migration distance of melanoma cells was significantly increased and the relative number of invasive cells was significantly decreased after curcumin treatment,and the differences were statistically significant(P<0.001;P<0.05).Western blot showed that the expression levels of Cyclin D1,Cyclin A2 and FoxM1 were decreased after curcumin treatment(P<0.001,P<0.05).Overexpression of FoxM1 up-regulated expression of NEDD4,knock-down of FoxM1 down-regulated NEDD4(P<0.001).Compared with the control group,the level ofβ-catenin in the nucleus was significantly decreased,while the level ofβ-catenin in the cytoplasm was significantly increased after curcumin treatment(P<0.001).Conclusion Curcumin inhibits the proliferation,migration and invasion of melanoma cells through down-regulating FoxM1,NEDD4 andβ-catenin.

关 键 词：姜黄素 黑素瘤 神经前体细胞表达发育下调蛋白4 叉头框蛋白M1 Β-连环蛋白 

分 类 号：R285[医药卫生—中药学]