罗氟司特对肝癌细胞WNT/β-连环蛋白信号的影响  

Effect of Roflumilast on WNT/β-catenin Signaling in Hepatocellular Carcinoma Cells

作　　者：彭俊凤[1] 郭平学 杨自华[2] Peng Junfeng;Guo Pingxue;Yang Zihua(Experimental Training Center of Shaanxi Energy Institute,Xi＇an 710613,China;Department of Hepatobiliary Surgery,the Second Affiliated Hospital of Xi＇an Medical University)

机构地区：[1]陕西能源职业技术学院实验实训中心,西安710613 [2]西安医学院第二附属医院肝胆外科

出　　处：《中国药师》2018年第11期1936-1941,共6页China Pharmacist

摘　　要：目的:探讨磷酸二酯酶4(PED4)抑制剂罗氟司特对肝癌细胞WNT/β-连环蛋白信号的影响。方法:不同浓度罗氟司特(10、20、40μmol·L^(-1))处理肝癌细胞HepG2,以未用罗氟司特处理的作为对照组,CKK-8法检测HepG2细胞增殖情况;流式细胞术检测HepG2细胞凋亡情况;Transwell法检测HepG2细胞的迁移和侵染能力;实时荧光定量PCR(q-PCR)和蛋白印记(WB)法检测细胞中相关蛋白GSK3β、p-GSK3β(ser9)、β-catenin、p-β-catenin和cyclin D1的mRNA水平和蛋白表达水平。结果:与对照组相比,10、20、40μmol·L^(-1)罗氟司特处理后,肝癌细胞HepG2的增殖能力显著降低,凋亡能力显著增强,迁移能力和侵袭能力显著降低,GSK3β、β-catenin和cyclin D1的mRNA表达水平显著降低,GSK3β、β-catenin、p-GSK3β(ser9)、p-β-catenin和cyclin D1的蛋白水平明显下降(P<0.05)。结论:磷酸二酯酶4(PED4)抑制剂能够促进肝癌细胞的凋亡,抑制肝癌细胞的增殖、迁移和侵袭。这一作用可能是通过抑制肝癌细胞HepG2中WNT/β-连环蛋白信号通路相关因子的表达实现的。Objective： To investigate the effect of phosphodiesterase 4 inhibitor roflumilast on WNT/beta-eatenin signaling in hepatocellular carcinoma cells. Methods： The HepG2 ceils were treated with different concentrations of roflumilast （10, 20 and 40 μmol·L^-1） and the inhibitory effects on HepG2 cells were examined. The proliferation of the HepG2 cells was detected by CKK-8 assay and the apoptosis was assayed by flow cytometry. The Transwell assay was conducted to evaluate the migration and invasion of HepG2 cells. Furthermore, quantitative real-time PCR was used to detect the mRNA level of GSK3β, β-catenin and cyclin D1 in the cells. And then, the protein levels of GSK3β, β-catenin, p-GSK3β （ser9） , p-β-catenin and cyclin DI were observed by using Western blot. Results： Compared with that in the control group, the proliferation of HepG2 cells was significantly decreased, and the apoptosis was significantly enhanced after the treatment of various concentrations of phosphodiesterase 4 inhibitor （P 〈 0.05 ）. And the phos- phodiesterase 4 inhibitor reduced the migration and invasion capability of HepG2 cells in a concentration-dependent manner （ P 〈 0.05）. Furthermore, the phosphodiesterase 4 inhibitor reduced the mRNA levels of GSK3β, β-catenin and cyclin D1 in the cells （P 〈 0.05）. In addition, the protein levels of GSK3β, β-catenin, p-GSK3β （ser9）, p-β-catenin and cyclin D1 were decreased （P 〈 0.05 ）. Conclusion ： Phosphodiesterase-4 inhibitor can promote the apoptosis of hepatocellular carcinoma cells and inhibit the prolifer- ation, migration and invasion of the cells. And the WNT/β- catenin signaling pathway may play an important role during the process.

关 键 词：罗氟司特 肝细胞癌 WNT/β-连环蛋白信号 

分 类 号：R965.1[医药卫生—药理学]