维罗非尼联合西达苯胺促进继发性急性髓系白血病细胞衰老  

Vemurafenib combined with chidamide promotes senescence of secondary acute myeloid leukemia cells

作　　者：周巧[1,2] 梁思敏[1] 蔡铎 向文琼 王利[1] ZHOU Qiao;IIANG Simin;CAI Duo;XIANG Wenqiong;WANG li(Department of Hematology,the First Affiliated Hospital of Chongqing Medical University,Chongqing,400016,China;Key Laboratory of Translational Medicine for Major Metabolic Diseases,the First Affiliated Hospital of Chongqing Medical University,Chongqing,400016,China)

机构地区：[1]重庆医科大学附属第一医院血液内科,重庆400016 [2]重庆医科大学附属第一医院重大代谢性疾病转化医学重点实验室,重庆400016

出　　处：《陆军军医大学学报》2023年第16期1671-1681,共11页Journal of Army Medical University

基　　金：重庆市渝中区科学技术局科学项目(20190121)。

摘　　要：目的探索维罗非尼(vemurafenib,VEM)联合西达苯胺(chidamide,CDM)方案对继发性急性髓系白血病(secondary acute myeloid leukemia,sAML)细胞衰老的影响。方法以sAML细胞系为研究对象(n=3),实验分为对照组、VEM单药组、CDM单药组、VEM+CDM组。CCK-8实验检测sAML细胞增殖抑制率;免疫荧光技术检测sAML细胞衰老发生;流式细胞术检测sAML细胞周期分布及凋亡发生;Swiss Target Predict、TargetNe、SEA及PharmMapper等4个网络药理学数据库预测VEM联合CDM的可能靶点和信号通路;RT-qPCR实验分析及验证VEM联合CDM的可能机制。结果VEM以时间剂量依赖性抑制sAML细胞增殖,VEM的半数抑制浓度(IC_(50))在SKM-1和MOLM-13细胞中分别为(24.15±1.18)μmol/L(P<0.0001)和(11.30±0.38)μmol/L(P<0.0001),VEM(SKM-1:1/3 IC_(50)-VEM,MOLM-13:1/2 IC_(50)-VEM)联合CDM协同抑制sAML细胞增殖(P<0.01);VEM联合CDM促进sAML细胞衰老相关标志物TRF2和Lamin B1表达;VEM联合CDM可使sAML细胞周期停滞于G_0/G_1期(P<0.05),并促进sAML细胞凋亡发生(P<0.05)。同时,网络药理学及RT-qPCR实验分析并验证VEM协同CDM通过抑制PI3K-AKT及MAPK信号通路促进sAML肿瘤细胞衰老发生(P<0.05)。结论VEM联合CDM通过下调PI3K-AKT及MAPK信号通路促进sAML细胞衰老发生、抑制sAML细胞增殖及促进sAML细胞凋亡,提示VEM联合CDM方案可能是sAML患者新的治疗方案。Objective To explore the effect of vemurafenib(VEM)combined with chidamide(CDM)on cellular senescence in secondary acute myeloid leukemia(sAML).Methods sAML cell lines.SKM-1 and MOLM-13 were divided into control group(sAML cells),VEM group(sAML cells treated with VEM),CDM group(sAML cells treated with CDM),and COM group(sAML cells treated with VEM combined with CDM)(n=3).CCK-8 assay was used to measure the viability of sAML cells.Immunofluorescence assay was used to verified the cellular senescence of sAML cells.Flow cytometry was used to detect cell cycle and apoptosis of sAML cells.Network pharmacology and RT-qPCR were used to analyzxe and verify the mechanism of VEM combined with CDM.Results VEM inhibited the prolferation of sAML cells lines in a time-and dose-dependent manner(P<0.05),the half-maximal inhibitory concentration(IC_(50))of VEM on SKM-1 and MOLM-13 cells was 24.15+1.18 and 11.30+0.38μmol/L(P<0.0001),respectively.VEM(SKM-1:1/3 ICso-VEM,MOLM-13:1/2 IC_(50)-VEM)combined with CDM synergi stically promoted sAML cellular senescence by upregulating TRF2 and Lamin BI,characterized cycle stagnation in G/G phase(P<0.05),inhibiting cell proliferation(P<0.01)and promoting cell apoptosis(P<0.05).Swiss Target Predict,TargetNe,SEA and PharmMapper discovered the core targets and signaling pathways of VEM colaborating with CDM.RT-qPCR verified that VEM collaborated with CDM to promote senescence of sAML cells through inhibiting PI3K-AKT and MAPK signaling pathways(P<0.05).Conclusion VEM combined with CDM can induce cellular senescence of sAML cells by downregulating PI3K-AKT and MAPK signaling pathways,inhibiting proliferation and promoting apoptosis of sAML cells.VEM combined with CDM might be a new regime for patients with sAML.

关 键 词：继发性急性髓系白血病 维罗非尼 西达苯胺 细胞衰老 

分 类 号：R73-361[医药卫生—肿瘤] R733.71[医药卫生—临床医学] R979.1