鼠尾草酸通过减少β-catenin表达抑制人黑素瘤A375细胞增殖  

Carnosic Acid Inhibits the Proliferation of Human Melanoma A375 Cells Through Attenuating β-catenin Expression

作　　者：王倩[1] 万品文[2] 李红文[3] WANG Qian;WAN Pinwen;LI Hongwen(Department of Dermatology,Nanyang Central Hospital,Nanyang 473000,China;Department of General Surgery,Nanyang Central Hospital,Nanyang 473000,China;Department of Dermatology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]南阳市中心医院皮肤科,河南南阳473000 [2]南阳市中心医院普外科,河南南阳473000 [3]郑州大学第一附属医院皮肤科,河南郑州450052

出　　处：《中国皮肤性病学杂志》2018年第10期1113-1119,共7页The Chinese Journal of Dermatovenereology

摘　　要：目的研究鼠尾草酸(CA)对人黑素瘤A375细胞增殖的影响并初步探讨其作用机制,为寻找新的黑素瘤临床治疗药物奠定一定的理论基础。方法 A375细胞传代培养后分为正常组、10μmol/L CA组(10μmol/L CA处理24h)、20μmol/L CA组、50μmol/L CA组,FH535 (β-catenin抑制剂)组(20μmol/L FH535处理24h)。MTT法检测细胞抑制情况;平板细胞克隆形成实验检测克隆形成情况;流式细胞仪检测细胞周期以及细胞凋亡变化; RT-PCR法检测A375细胞中β-catenin、细胞周期蛋白D1(CyclinD1)、淋巴增强因子1(LEF1)、原癌基因(C-myc) mRNA的表达; Western blot检测A375细胞中β-catenin、CyclinD1、LEF1、C-myc蛋白表达。结果与正常组相比,CA各组、FH535组细胞抑制率增加,CA各组呈时间、剂量依赖性增加(P <0. 05); CA各组、FH535组克隆形成数量均降低,CA各组克隆形成数量呈剂量依赖性降低(P <0. 05)。流式细胞仪检测显示与正常组相比,CA各组、FH535组G1期细胞比例升高,S期细胞比例降低,CA各组G1期细胞比例呈剂量依赖性升高,S期细胞比例呈剂量依赖性降低(P <0. 05); CA各组、FH535组细胞凋亡率升高,CA各组凋亡率呈剂量依赖性升高(P <0. 05)。与正常组相比,CA各组、FH535组β-catenin、CyclinD1、LEF1、C-myc mRNA、蛋白表达降低,CA各组呈剂量依赖性降低(P <0. 05)。结论 CA可能通过抑制β-catenin的表达,引起细胞周期G1期阻滞,进而抑制人黑素瘤A375细胞增殖。Objective To study the effect of carnosic acid（CA） on the proliferation of human melanoma A375 cells and related mechanisms,and to provide certain theoretical foundations for searching new drugs in the treatment of melanoma. Methods A375 cells were divided into normal group,10μmol/L CA group（10μmol/L CA treated 24 h）,20μmol/L CA group,50μmol/L CA group,FH535（β-catenin inhibitor） group（20μmol/L FH535 treated for 24 h） after subculture. MTT assay and clone formation assay were used to detect cell viability and proliferation. Cell cycle and apoptosiswere determined by flow cytometry. RT-PCR was used to detect the mRNA expressions of β-catenin,Cyclin D1（CyclinD1）,lymphatic enhancement factor 1（LEF1）,and proto oncogene（C-myc） in A375 cells. Western blot was applied to detect the protein expressions of β-catenin,CyclinD1,LEF1 and C-myc in A375 cells. Results In comparison to the normal group,cell inhibition rate was increased in a time and dose dependent way in the CA groups and FH535 group（P〉0. 05）. The numbers of colonies in each group of CA and FH535 group decreased in a dose-dependent manner（P〉0. 05）. Flow cytometry showed that the proportions of G1 phase cells in CA groups and FH535 group were higher than those in normal group,while the proportion of S phase cells decreased（P〉0. 05）; The apoptotic rates of CA groups and FH535 group increased,the apoptotic rates of CA groups increased in a dose-dependent manner（P〉0. 05）. The mRNA and protein expressions of β-catenin,CyclinD1,LEF1,C-myc in CA groups and FH535 group attenuated in a dose-dependent manner（P〉0. 05）. Conclusion CA might block the G1 phase of cell cycle by attenuating the expression of β-catenin,thus inhibit the proliferation of human melanoma A375 cells.

关 键 词：鼠尾草酸 人黑素瘤A375细胞 Β-链蛋白 作用机制 

分 类 号：R739.5[医药卫生—肿瘤]