新型磷酸二酯酶4抑制剂ZL-n-91对骨肉瘤U2OS细胞增殖的抑制作用  

The inhibitory effect of a novel phosphodiesterase 4 inhibitor ZL-n-91 on proliferation of osteosarcoma U2OS cells

作　　者：许丽君 王学鹏 林伟杰 梁龙铭 赫玉杰 赵正刚 李芳红 周素瑾 赵子建 XU Li-jun;Wang Xue-peng;LIN Wei-jie;LIANG Long-ming;HE Yu-jie;ZHAO Zheng-gang;LI Fang-hong;ZHOU Su-jin;ZHAO Zi-jian(School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Guangzhou 510006, China)

机构地区：[1]广东工业大学生物医药学院,广东广州510006

出　　处：《中国药理学通报》2022年第2期233-238,共6页Chinese Pharmacological Bulletin

基　　金：国家重点研发计划(No 2018YFA0800603);广东省重点领域研发计划“新药创制”专项(No 2019B020201015);广东省创新团队(No 2016ZT06Y432);广东省高校高水平科研项目(No 20ZK0230)。

摘　　要：目的研究高选择性的新型磷酸二酯酶4抑制剂ZL-n-91对骨肉瘤的抗癌作用。方法CCK-8法检测不同浓度(0、20、40、80、160、240、320、400、480μmol·L^(-1))和不同干预时间(0、24、48、72、96 h)的ZL-n-91对U2OS细胞的增殖抑制作用;平板克隆形成实验检测ZL-n-91对U2OS细胞克隆形成能力的影响;流式细胞术检测细胞周期以及凋亡的影响;Western blot检测抗凋亡蛋白Bcl-2、周期蛋白CDK2、结果ZL-n-91明显抑制U2OS细胞的增殖,呈时间及剂量依赖性(P<0.05),其半数抑制浓度IC_(50)为174.1μmol·L^(-1);ZL-n-91明显抑制U2OS细胞的克隆形成能力(P<0.01);ZL-n-91影响U2OS细胞的周期进程,将U2OS细胞阻滞在G_(0)/G_(1)期,并明显促进细胞凋亡(P<0.01);Western blot结果显示ZL-n-91明显下调U2OS细胞中Bcl-2、CDK2、CDK4、CyclinD1、CyclinE1蛋白的表达(P<0.05)。结论新型选择性磷酸二酯酶4抑制剂ZL-n-91显著抑制骨肉瘤细胞U2OS的增殖,促进细胞的周期阻滞和凋亡,可能是治疗骨肉瘤的潜在药物。Aim To explore the anti-cancer effects of ZL-n-91,a novel and highly selective phosphodiesterase 4 inhibitor,on the osteosarcoma U2OS cells.Methods CCK-8 assay was used to detect the inhibitory effect of ZL-n-91 with different concentrations(0,20,40,80,160,240,320,400,480μmol·L^(-1))and different intervention time(0,24,48,72,96 h)on the proliferation of U2OS cells.Tablet clone forming experiment was used to detect the effect of ZL-n-91 on the clonality of U2OS cells.Flow cytometry was used to detect the cell apoptosis and cell cycle distribution.Western blot was employed to detect the expression of Bcl-2,CDK2,CDK4,CyclinD1,CyclinE1 protein.Results The inhibitory rate of ZL-n-91 on U2OS cells was concentration-and time-dependent(P<0.05),and its half inhibition rate IC_(50) was 174.1μmol·L^(-1).ZL-n-91 significantly inhibited the clonality of U2OS cells(P<0.01).ZL-n-91 significantly induced cell apoptosis,and caused cell cycle arrest at G_(0)/G_(1) phase in U2OS cells(P<0.01).The results of Western blot showed that ZL-n-91 significantly down-regulated the expression of Bcl-2,CDK2,CDK4,CyclinD1,CyclinE1 proteins in U2OS cells(P<0.05).Conclusions The novel selective phosphodiesterase 4 inhibitor,ZL-n-91,can significantly inhibit the proliferation of osteosarcoma U2OS cells with induction of cell cycle arrest and cell apoptosis,and may become a potential anti-cancer agent.

关 键 词：骨肉瘤 ZL-n-91 U2OS细胞 增殖 凋亡 周期 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R329.28[医药卫生—基础医学]