新型小分子化合物cyy-287联合CDK抑制剂roscovitine抑制SBC-2细胞生长的机制  

The inhibitory mechanism of a novel small molecular compound cyy-287 combined with CDK inhibitor roscovitine in SBC-2 cell growth

作　　者：郑淑文 黄慧静 陈国 麻浩群 钱建畅 张倩雯 ZHENG Shuwen;HUANG Huijing;CHEN Guo;MA Haoqun;QIAN Jianchang;ZHANG Qianwen(School of Pharmaceutical Sciences,Wenzhou Medical University,Wenzhou 325035,China)

机构地区：[1]温州医科大学药学院,浙江温州325035

出　　处：《温州医科大学学报》2023年第2期109-114,共6页Journal of Wenzhou Medical University

基　　金：浙江省自然科学基金项目(LQ21H310007);浙江省大学生科技创新活动计划(新苗人才计划)项目(2022R413A008);温州市基础性科研项目(2020Y1214,Y20220200)。

摘　　要：目的:探讨新型小分子化合物cyy-287联合细胞周期蛋白依赖性激酶(CDK)抑制剂roscovitine抑制小细胞肺癌细胞SBC-2生长的机制。方法:取对数生长期的SBC-2细胞,通过MTS检测cyy-287和顺铂(DDP)的IC50值,以及cyy-287和roscovitine联合使用的联合指数(CI)。cyy-287与roscovitine联用抗肿瘤活性实验分组:DMSO(对照)组、roscovitine(20μmol/L)组、cyy-287(1.5μmol/L)组、cyy-287+roscovitine组。通过克隆形成实验检测各组药物处理后肿瘤细胞的生长和增殖情况;流式细胞术测定细胞周期分布和凋亡比例的变化情况。采用Western blot法检测各组CL-PARP的蛋白表达水平,以及AKT的磷酸化水平。结果:cyy-287对SBC-2的IC50值为(1.77±0.10)μmol/L,与roscovitine联合使用的CI<1,说明两药联合使用存在协同作用。与cyy-287组比,cyy-287+roscovitine组能够显著抑制SBC-2的生长和克隆形成(P<0.001),且凋亡细胞比例明显增加(P<0.001);凋亡相关CL-PARP的蛋白表达水平显著上升,P-AKT表达水平下降(P<0.05)。结论:cyy-287与roscovitine联用具有协同的抗肿瘤作用,可有效抑制SBC-2细胞生长并诱导其凋亡,其潜在机制主要是通过抑制AKT的磷酸化,抑制细胞存活。Objective:To explore the inhibitory effect of a new small molecule compound cyy-287,combined with cyclin dependent kinase(CDK)inhibitor roscovitine on the small cell lung cancer cell line SBC-2 and its mechanism.Methods:SBC-2 cells in logarithmic growth period were taken.The IC50values of cyy-287 and the combination index(CI)of cyy-287 and roscovitine were detected by MTS.Experimental groups of cyy-287 in combination with roscovitine for antitumor activity studies were as follows:DMSO(control)group;roscovitine(20μmol/L)group;cyy-287(1.5μmol/L)group and cyy-287+roscovitine group.The proliferation of tumor cells after drug treatment in each group was detected by clone formation experiment.The distribution of cell cycle and the proportion of apoptosis were measured by flow cytometry.The protein expression level of CLPARP and the phosphorylation level of AKT in each treatment group were detected by Western blot.Results:The IC50of cyy-287 in SBC-2 was(1.77±0.10)μmol/L.The CI of cyy-287 and roscovitine was less than 1,which means that cyy-287 combined with rescovitine has synergistic effect on SBC-2 cells.Compared with cyy-287alone treatment group,a combination of the two drugs could significantly inhibit the growth and clonogenesis of SBC-2 cells(P<0.001),and the proportion of apoptotic cells increased significantly(P<0.001).The protein expression level of apoptosis related CL-PARP increased significantly,and the expression level of P-AKT decreased(P<0.05).Conclusion:The combination of cyy-287 and roscovitine has a synergistic antitumor effect,which can effectively inhibit the growth of SBC-2 cells and induced their apoptosis.Its underlying mechanism is mainly suppressing the cell survival by inhibiting the phosphorylation of AKT.

关 键 词：cyy-287 ROSCOVITINE 小细胞肺癌 细胞凋亡 

分 类 号：R780.1[医药卫生—口腔医学]