硼替佐米联合AZD-1775对人多发性骨髓瘤细胞株凋亡的影响  

Effect of bortezomib combined with AZD-1775 on apoptosis of human multiple myeloma cell line

作　　者：马莉 李增慧 荣芳 王建宾 张年萍 MA Li;LI Zenghui;RONG Fang;WANG Jianbin;ZHANG Nianping(Department of Internal Medicine,Medical College of Datong University,Datong 037009,China;Department of Orthopedics,Datong Third People’s Hospital)

机构地区：[1]山西大同大学医学院内科教研室,大同037009 [2]大同市第三人民医院骨科

出　　处：《山西医科大学学报》2023年第1期48-52,共5页Journal of Shanxi Medical University

基　　金：山西大同大学校级重点建设学科项目(3)。

摘　　要：目的探讨硼替佐米联合WEE1激酶抑制剂AZD-1775对人多发性骨髓瘤(MM)细胞凋亡的影响。方法培养RPMI8226和U266细胞,逆转录PCR(RT-PCR)检测WEE1 mRNA表达。采用随机数字表法将两种细胞分为对照组、AZD-1775组、硼替佐米组和联合处理组。AZD-1775组细胞加入AZD-1775(10μmol/L)孵育24 h;硼替佐米组细胞加入硼替佐米(20 nmol/L)孵育24 h;联合处理组细胞先加入硼替佐米(20 nmol/L)孵育24 h,再加入AZD-1775(10μmol/L)孵育24 h。采用肿瘤细胞成球实验检测不同处理对RPMI8226和U266细胞成球率影响;采用流式细胞术检测不同处理对RPMI8226和U266细胞凋亡率影响;采用Westen blot检测不同处理对RPMI8226细胞Bcl-2和Caspase-3蛋白表达影响。结果RT-PCR检测显示,RPMI822和U266细胞均存在WEE1 mRNA表达。与对照组相比,AZD-1775组、硼替佐米组和联合处理组细胞球形成率均降低(P<0.05),而联合处理组细胞球形成率低于AZD-1775组和硼替佐米组(P<0.05)。与对照组相比,AZD-1775组、硼替佐米组和联合处理组RPMI-8226和U266细胞总凋亡率均增高(P<0.05),而联合处理组高于AZD-1775组和硼替佐米组(P<0.05)。此外,与对照组相比,AZD-1775组、硼替佐米组和联合处理组Bcl-2蛋白水平均降低,Caspase-3蛋白水平均增加(P<0.05),而联合处理组Bcl-2蛋白水平低于AZD-1775组和硼替佐米组,Caspase-3蛋白水平高于AZD-1775组和硼替佐米组(P<0.05)。结论硼替佐米联合AZD-1775可更有效诱导RPMI8226及U266细胞凋亡,其机制可能与Bcl-2表达下调,Caspase-3表达增加有关。Objective To investigate the effect of bortezomib combined with WEE1 kinase inhibitor AZD-1775 on apoptosis of human multiple myeloma(MM)cells.Methods RPMI8226 and U266 cells were cultured and randomized into control group,AZD-1775 group,bortezomib group and combined treatment group.RPMI8226 and U266 cells were incubated with AZD-1775(10μmol/L)for 24 h in AZD-1775 group,and bortezomib(20 nmol/L)for 24 h in bortezomib group.In combined treatment group,RPMI8226 and U266 cells were incubated with bortezomib(20 nmol/L)for 24 h,and then AZD-1775(10μmol/L)for 24 h.The sphere forming rates of RPMI8226 and U266 cells were determined by tumor cell pellet-forming assay.The apoptosis rates of RPMI8226 and U266 cells were detected by flow cytometry.The effects of different treatments on the expression of Bcl-2 and Caspase-3 protein in RPMI8226 cells were detected by Westen blot.Results Reverse transcription PCR(RT-PCR)results showed that WEE1 mRNA was expressed in both RPMI822 and U266 cells.The sphere forming rate was lower in AZD-1775 group,bortezomib group and combined treatment group than in control group(P<0.05),and the sphere forming rate in combined treatment group was lower than that in AZD-1775 and bortezomib group(P<0.05).The total apoptosis rates of RPMI-8226 and U266 cells in AZD-1775 group,bortezomib group and combined treatment group were higher than in control group(P<0.05),and also higher in combined treatment group than in AZD-1775 and bor-tezomib group(P<0.05).Compared with control group,the level of Bcl-2 protein was reduced and the level of Caspase-3 protein was increased in AZD-1775 group,bortezomib group and combined treatment group(P<0.05),moreover,compared with AZD-1775 group and bortezomib group,the level of Bcl-2 protein in combined treatment group was decreased and the level of Caspase-3 protein was increased(P<0.05).Conclusion Bortezomib combined with AZD-1775 could more effectively induce the apoptosis of RPMI8226 and U266 cells,which may be related to the down-regulation of Bcl-2 expression an

关 键 词：硼替佐米 AZD-1775 凋亡 RPMI8226细胞 U266细胞 

分 类 号：R733[医药卫生—肿瘤]