新型Ⅰ类和Ⅱb类选择性HDAC抑制剂甲磺酸普依司他治疗弥漫大B细胞淋巴瘤的体外药效学和作用机制研究  

In vitro pharmacodynamic studies of novel classⅠandⅡb selective histone deacetylase inhibitor purinostat mesylate in the treatment of diffuse large B-cell lymphoma and its mechanism

作　　者：王婕[1] 赵艾琳 李赫[1] 杨林玉 陈俐娟[2] 牛挺[1] Wang Jie;Zhao Ailin;Li He;Yang Linyu;Chen Lijuan;Niu Ting(Department of Hematology,West China Hospital,Sichuan University,Chengdu 610041,China;State Key Laboratory of Biotherapy and Cancer Center,West China Hospital,Sichuan University,Chengdu 610041,China)

机构地区：[1]四川大学华西医院血液内科,成都610041 [2]四川大学生物治疗国家重点实验室,成都610041

出　　处：《中华血液学杂志》2022年第9期753-759,共7页Chinese Journal of Hematology

基　　金：四川大学华西医院临床研究孵化项目(19HXFH030);四川大学华西医院学科卓越发展1.3.5工程项目(ZYJC21007)。

摘　　要：目的探究新型Ⅰ类和Ⅱb类选择性组蛋白去乙酰化酶(HDAC)抑制剂甲磺酸普依司他(PM)对弥漫大B细胞淋巴瘤的体外抑制活性及作用机制。方法 MTT法检测PM对细胞增殖的影响, 流式细胞术检测PM对细胞周期、细胞凋亡的影响, Western blot法检测HDAC底物乙酰化水平、细胞周期蛋白、凋亡相关蛋白和癌基因蛋白的表达。结果 PM对淋巴瘤细胞株SUDHL-4和SUDHL-6的增殖具有明显抑制作用, 能够上调HDAC底物H3、H4和α-tubulin乙酰化水平。在细胞周期实验中, PM可诱导SUDHL-4、SUDHL-6细胞G0/G1期阻滞, Western blot显示PM能显著下调细胞周期蛋白依赖性激酶Cdk2、Cdk4、Cdk6及细胞周期蛋白Cyclin D1、Cyclin E的表达, 上调CDK抑制蛋白p21的表达。在细胞凋亡实验中, PM可诱导SUDHL-4和SUDHL-6细胞凋亡, Western blot显示PM通过激活Caspase3激酶和影响抗凋亡蛋白Bcl-2促进细胞内源性凋亡。此外, PM也能够下调癌基因标志蛋白MYC、IKZF1、IKZF3的表达。结论 PM在体外对弥漫大B细胞淋巴瘤包括双打击淋巴瘤具有高效的生物活性, 为PM应用于临床治疗提供了有价值的实验依据。Objective To investigate the in vitro inhibitory activity of a novel classⅠandⅡb selective histone deacetylase(HDAC)inhibitor,purinostat mesylate(PM),in diffuse large B-cell lymphoma and its mechanism.Methods The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide method was used to detect the effect of PM on cell proliferation.The effects of PM on cell cycle and apoptosis were detected by flow cytometry.The acetylation levels of HDAC substrate,cell cycle protein,apoptosis-related protein,and oncogene protein expression were detected by Western blot.Results PM significantly inhibited the proliferation of lymphoma SUDHL-4 and SUDHL-6 cells and increased the acetylation levels of HDAC substrates H3,H4,andα-tubulin.In cell cycle experiments,PM induced G_(0)/G_(1) phase arrest in SUDHL-4 and SUDHL-6 cells.Western blot experiment showed that PM could significantly downregulate the expression of cyclin-dependent kinases Cdk2,Cdk4,Cdk6,cyclin D1,and cyclin E and upregulate the expression of CDK inhibitor protein p21.In the apoptosis experiment,PM could induce the apoptosis of SUDHL-4 and SUDHL-6 cells.Western blot experiment demonstrated that PM promoted endogenous apoptosis by activating caspase-3 kinase and affecting antiapoptotic protein Bcl-2.In addition,PM could downregulate the expression of oncogene marker proteins MYC,IKZF1,and IKZF3.Conclusion PM has an efficient biological activity in vitro for diffuse large B-cell lymphoma,including double-hit lymphoma,and provides valuable experimental evidence for PM in clinical treatment.

关 键 词：组蛋白脱乙酰基酶抑制剂 淋巴瘤 大B细胞 弥漫性 体外药效学 作用机制 

分 类 号：R733.1[医药卫生—肿瘤]