新型磷酸二酯酶4抑制剂ZL-n-91对人神经胶质瘤细胞增殖、周期及凋亡的影响

Effect of a novel phosphodiesterase 4 inhibitor ZL-n-91 on proliferation,cell cycle and apoptosis of human glioma U87 cells

目的探讨新型磷酸二酯酶4抑制剂ZL-n-91对人神经胶质瘤U87细胞增殖、周期及凋亡的影响。方法体外实验,CCK-8法检测不同浓度的ZL-n-91作用U87细胞48 h后细胞增殖抑制情况;流式细胞术检测U87细胞周期和凋亡变化;Western blot检测细胞周期蛋白CDK2、CDK4、Cyclin D1及凋亡蛋白Bax和Bcl-2的表达;体内实验,构建U87皮下移植瘤模型,检测5 mg·kg^(-1) ZL-n-91对神经胶质瘤的治疗作用。结果ZL-n-91呈浓度依赖性抑制U87细胞增殖(P<0.01),IC50值为274.5μmol·L^(-1);与对照组相比,经ZL-n-91处理后U87细胞出现明显的G0/G1期阻滞(P<0.01),并呈现剂量依赖性诱导细胞凋亡(P<0.05);Western blot结果表明,ZL-n-91降低了胶质瘤细胞CDK2、CDK4、Cyclin D1、Bcl-2蛋白的表达(P<0.05);体内实验表明,ZL-n-91显著抑制U87皮下移植瘤的生长(P<0.05)。结论新型PDE4抑制剂ZL-n-91显著抑制神经胶质瘤U87细胞的增殖,将细胞阻滞在G0/G1期,可能通过Bcl-2/Bax途径诱导细胞凋亡。

Aim To investigate the effect of ZL-n-91,a novel phosphodiesterase 4(PDE4)inhibitor,on proliferation,cell cycle and apoptosis of human glioma U87 cells.Methods In vitro the different concentrations of ZL-n-91 were set up to evaluate the proliferation of U87 cells by CCK-8 assay.The cell apoptosis and cell cycle distribution were examined by flow cytometry.The expression of CDK2,CDK4,cyclin D1 and apoptosis-related protein Bax and Bcl-2 proteins were detected by Western blot.A subcutaneous transplanted tumor model of U87 in nude mice was established for the in vivo experiment using a dosage of 5mg·kg^(-1) of ZL-n-91.Results ZL-n-91 significantly inhibited the proliferation of U87 cells with concentration dependence,and the IC50 value was 274.5μmol·L^(-1).In addition,ZL-n-91 induced G0/G1 cell cycle arrest in U87 cancer cells(P<0.0l).Meanwhile,ZL-n-91 treatment induced apoptosis in a dose-dependent manner.Western blot analysis demonstrated that ZL-n-91 down-regulated the expression of CDK2,CDK4,cyclin D1 and Bcl-2(P<0.05).Furthermore,ZL-n-91 significantly inhibited the growth of transplanted U87-derived tumor in the nude mice.Conclusions ZL-n-91,a novel PDE4 inhibitor,can significantly inhibit the proliferation of U87 cells,promote cell arrest in G0/G1 phase,and probably induce cell apoptosis through the Bcl-2/Bax pathway.