橙花叔醇通过Wnt-β-catenin通路调控黑色素瘤细胞的恶性生物学行为  

Nerolidol inhibits malignant biological behaviors of melanoma cells by regulating the Wnt-β-catenin pathway

作　　者：刘曌 王建澍[1] 薛金旭 朱彦奇 李晶[1] LIU Zhao;WANG Jianshu;XUE Jinxu;ZHU Yanqi;LI Jing(Second Department of Bone and Soft Tissue Oncology,Gansu Provincial Cancer Hospital,Lanzhou 730050,Gansu,China)

机构地区：[1]甘肃省肿瘤医院骨与软组织肿瘤二科,甘肃兰州730050

出　　处：《中国肿瘤生物治疗杂志》2023年第3期204-210,共7页Chinese Journal of Cancer Biotherapy

基　　金：甘肃省卫生行业科研项目(No.GSWSKY-2019-30)。

摘　　要：目的:探讨橙花叔醇通过Wnt-β-catenin通路抑制黑色素瘤A-375和WM-115细胞恶性生物学行为的分子机制。方法:体外培养黑色素瘤细胞A-375和WM-115,用不同浓度的橙花叔醇处理,采用SRB法和克隆形成实验、FCM术、Transwell实验和细胞划痕实验、DCFH-DA染色法、qPCR和WB法分别检测橙花叔醇对A-375和WM-115细胞的增殖能力、细胞周期和凋亡、迁移能力、活性氧(ROS)水平和Wnt-β-catenin通路及其下游相关基因和相关蛋白表达的影响。利用ULCAN和GEPIA2数据库分析黑色素瘤中Wnt-β-catenin通路的激活与患者预后的关系。结果:与对照组比较,橙花叔醇处理组A-375和WM-115细胞的增殖能力受明显抑制(均P<0.01)、细胞周期阻滞于G2/M期(P<0.05或P<0.01)、细胞凋亡率增加(均P<0.01)、迁移能力降低(P<0.05或P<0.01)、ROS水平升高(均P<0.01)、Wnt-β-catenin通路被抑制而其下游基因和蛋白表达明显上调(均P<0.01)。数据库数据分析显示,WNT1基因高表达患者OS低于低表达患者(P<0.01)。结论:橙花叔醇通过上调A-375和WM-115细胞中ROS水平影响Wnt-β-catenin通路,从而抑制其恶性生物学行为;Wnt-β-catenin通路可能是黑色素瘤治疗的潜在靶点。Objective:To investigate the molecular mechanism by which nerolidol inhibits the malignant biological behavior of melanoma A-375 and WM-115 cells through the Wnt-β-catenin pathway.Methods:Melanoma A-375 and WM-115 cells were cultured in vitro and then treated with different concentrations of nerolidol.The effects of nerolidol on the proliferation,cell cycle and apoptosis,and migration of A-375 and WM-115 cells were analyzed by SRB and clonogenic assays,FCM,Transwell,and cell scratch assays,respectively.The levels of reactive oxygen species(ROS)in the cells were examined with DCFH-DA staining.The Wnt-β-catenin pathway and the expression levels of its related downstream genes were determined by qPCR and WB.The relationship between patient prognosis and the activation of Wnt-β-catenin pathway in melanoma was analyzed using the ULCAN and GEPIA2 databases.Results:Compared with the control group,the proliferation,migration,and cell cycle of A-375 and WM-115 cells in the nerolidol-treated group were significantly inhibited(all P<0.01),while the apoptosis was significantly increased(all P<0.01);the ROS level was increased(P<0.01),and the Wnt-β-catenin pathway was inhibited,while its downstream gene expression was significantly up-regulated(P<0.01 or P<0.01).Analysis of database data showed that OS was lower in patients with high WNT1 gene expression than in patients with low expression(P<0.01).Conclusions:Nerolidol affects the Wnt-β-catenin pathway by upregulating ROS levels in A-375 and WM-115 cells,thereby inhibiting their malignant biological behaviors.The Wnt-β-catenin pathway may be a potential target for the treatment of melanoma.

关 键 词：橙花叔醇 黑色素瘤 A-375细胞 WM-115细胞 Wnt-β-catenin通路 活性氧 

分 类 号：R739.5[医药卫生—肿瘤] R285[医药卫生—临床医学]