雷公藤甲素通过PTEN表达调控Wnt/β-catenin通路抑制黑色素瘤的机制研究  

Effect of Triptolide on the Regulation of Wnt/β-catenin Pathway on Inhibiting Melanoma by PTEN Expression

作　　者：张步鑫[1] 赵献敏[2] 成琼 李锡清[3] ZHANG Bu-xin;ZHAO Xian-min;CHENG Qiong;LIXi-qing(Dermatology Department in Hospital of Traditional Chinese Medicine of Henan Province,Zhengzhou 450002,China;Basic Medi- cal College,Henan University of Traditional Chinese Medicine,Zhengzhou 450046,China;Henan Provincial People's Hospital, Zhengzhou 450003,China)

机构地区：[1]河南省中医院皮肤科,河南郑州450002 [2]河南中医药大学基础医学院,河南郑州450046 [3]河南省人民医院,河南郑州450003

出　　处：《中药材》2018年第2期427-431,共5页Journal of Chinese Medicinal Materials

基　　金：国家自然科学基金(81603632)

摘　　要：目的:探讨雷公藤甲素的抗黑色素瘤作用是否与PTEN表达及Wnt/β-catenin通路相关。方法:实验分为5组:空白对照组、雷公藤甲素(20 nmol/L)组、β-catenin抑制剂(5μmol/L IWR-1-endo)组、雷公藤甲素+β-catenin抑制剂组、ATRA(100μmol/L)组。分别采用CCK8法、Annexin V-FITC/PI双染色法、实时定量PCR法和Western blot法检测各组A375细胞活力、凋亡率及PTEN、β-catenin、Bcl-2、Caspase-3、PCNA mRNA及蛋白的表达。结果:雷公藤甲素、β-catenin抑制剂和ARTA处理能抑制A375细胞的增殖并诱导其凋亡,在增强Caspase-3和PTEN表达的同时抑制β-catenin、Bcl-2、PCNA的表达。结论:雷公藤甲素能抑制黑色素瘤细胞A375的增殖并诱导其凋亡,该作用可能是通过增加PTEN的表达进而抑制Wnt/β-catenin通路的激活来实现。Objective :To investigate whether the anti-melanoma effect of Triptolide was related to PTEN expression and Wnt/β-catenin pathway.Methods:The experiment was divided into five groups,blank control group,Triptolide group(20nmol/L),β--catenin inhibitor group(5μmol/L IWR-1-endo),Triptolide+β-catenin inhibitor group(20nmol/L Triptolide+5μmo]/L IWR-1-endo),ATRA group(100μmo]/L ATRA).CCK8 assay,Annexin V-FITC/PI double staining,real-time quantitative PCR and Western blot were used to detect the cell vitality,apoptosis rate and the expressions of PTEN,β-catenin,Bcl-2,Caspase-3 and PCNA in A375 cells, respectively.Results:Triptolide,β-catenin inhibitor and ARTA could inhibit the proliferation of A375 cells and induce apoptosis.The expression of β-catenin,Bcl-2 and PCNA was also inhibited while enhancing the expression of Caspase-3 and PTEN.Conclusion:Triptolide can inhibit the proliferation of melanoma A375 cells and induce its apoptosis.This effect may be achieved by increasing the expression of PTEN and inhibiting the activation of Wnt/β-catenin pathway.

关 键 词：雷公藤甲素 PTEN WNT/Β-CATENIN通路 黑色素瘤 

分 类 号：R285.5[医药卫生—中药学]