CAJ | 학술논문

目的探讨磷脂酰肌醇3-激酶(PI3Kδ)抑制剂CAL-101对多发性骨髓瘤(multiple myeloma,MM)细胞增殖的影响及机制,为MM治疗提供新思路.方法以MM细胞系U266、RPMI8226及MM患者骨髓瘤原代细胞为研究对象,用不同浓度CAL-101处理后,采用MTT法结合CalcuSyn软件分析细胞增殖抑制率,并观察CAL-101与PCI-32765、异羟肟酸(SAHA)、硼替佐米(BTZ)联合对MM细胞增殖的影响;Western blot法分析蛋白激酶B(AKT)、细胞外调节蛋白激酶(ERK)、磷酸化AKT (p-AKT)、磷酸化ERK(p-ERK)和PI3Kδ表达水平.结果 15、20、25、30、40μmol/L CAL-101作用于U266细胞48 h,细胞增殖抑制率分别为(33.54± 1.23)％、(41.72±1.78)％、(53.67±2.01)％、(68.97±2.11)％和(79.25±1.92)％,具有明显的剂量依赖性(P＜0.05).对RPMI8226细胞的作用同对U266细胞的作用相似,亦呈明显的剂量依赖性.Western blot分析显示,U266、RPMI8226和MM原代细胞均可见AKT、ERK、p-AKT、p-ERK和PI3Kδ的表达;经CAL-101处理后,p-AKT和p-ERK的表达水平显著下调.CalcuSyn分析证实,对U266细胞,CAL-101与PCI-32765、SAHA、BTZ抑制细胞增殖具有协同作用,而对RPMI8226细胞,CAL-101与PCI-32765、BTZ抑制细胞增殖具有协同作用,协同指数小于1.结论 CAL-101可抑制MM细胞增殖.在MM细胞系及MM原代细胞均可见p-AKT、p-ERK、AKT、ERK和PI3Kδ的表达.CAL-101抑制MM细胞增殖的机制可能与下调p-AKT和p-ERK有关.CAL-101与MM一线治疗药物BTZ及新药PCI-32765、SAHA有显著协同抗MM效应.
목적 탐토린지선기순3-격매(PI3Kδ)억제제CAL-101대다발성골수류(multiple myeloma,MM)세포증식적영향급궤제,위MM치료제공신사로.방법 이MM세포계U266、RPMI8226급MM환자골수류원대세포위연구대상,용불동농도CAL-101처리후,채용MTT법결합CalcuSyn연건분석세포증식억제솔,병관찰CAL-101여PCI-32765、이간우산(SAHA)、붕체좌미(BTZ)연합대MM세포증식적영향;Western blot법분석단백격매B(AKT)、세포외조절단백격매(ERK)、린산화AKT (p-AKT)、린산화ERK(p-ERK)화PI3Kδ표체수평.결과 15、20、25、30、40μmol/L CAL-101작용우U266세포48 h,세포증식억제솔분별위(33.54± 1.23)％、(41.72±1.78)％、(53.67±2.01)％、(68.97±2.11)％화(79.25±1.92)％,구유명현적제량의뢰성(P＜0.05).대RPMI8226세포적작용동대U266세포적작용상사,역정명현적제량의뢰성.Western blot분석현시,U266、RPMI8226화MM원대세포균가견AKT、ERK、p-AKT、p-ERK화PI3Kδ적표체;경CAL-101처리후,p-AKT화p-ERK적표체수평현저하조.CalcuSyn분석증실,대U266세포,CAL-101여PCI-32765、SAHA、BTZ억제세포증식구유협동작용,이대RPMI8226세포,CAL-101여PCI-32765、BTZ억제세포증식구유협동작용,협동지수소우1.결론 CAL-101가억제MM세포증식.재MM세포계급MM원대세포균가견p-AKT、p-ERK、AKT、ERK화PI3Kδ적표체.CAL-101억제MM세포증식적궤제가능여하조p-AKT화p-ERK유관.CAL-101여MM일선치료약물BTZ급신약PCI-32765、SAHA유현저협동항MM효응.
Objective To investigate the proliferation inhibitory role and mechanism of PI3Kδ inhibitor CAL-101 on multiple myeloma (MM) cells,and to provide new therapeutic options for MM treatment.Methods MM cell lines U266 and RPMI8226 cells were treated with various concentrations of CAL-101.MTT assay and CalcuSyn software were performed to determine the inhibitory effect of CAL-101 and the synergistic effect with PCI-32765,SAHA (suberoylanilide hydroxamic acid),BTZ (Bortezomib) on MM cells.The protein expression level of p-AKT,p-ERK,AKT,ERK and PI3Kδ processed by CAL-101 were analyzed by Western blot.Results CAL-101 at concentration of 15,20,25,30 and 40 μmol/L could induce significant dose-dependent proliferation inhibition on U266 cells after treatment for 48 hours.The cell proliferation inhibition rates were (33.54±1.23)％,(41.72±1.78)％,(53.67±2.01)％,(68.97±2.11)％ and (79.25±1.92)％,respectively.Similar results were found in RPMI8226 cell line.Western blots showed high expression level of p-AKT,p-ERK,AKT,ERK and PI3Kδ in cell lines and MM primary cells,p-AKT and p-ERK protein expression levels were down-regulated significantly by CAL-101 treatment.Synergistic effect has been verified between CAL-101 and PCI-32765,SAHA and Bortezomib in U266 cell line,and PCI-32765,Bortezomib in RPMI8226 cell line with CI values less than 1.Conclusion CAL-101 could inhibit proliferation of MM cell lines.High levels of p-AKT,p-ERK,AKT,ERK and PI3Kδ protein expression were observed in both cell lines and primary cells.Down-regulation of p-AKT and p-ERK probably related with the mechanism of CAL-101 in MM cell proliferation inhibition.CAL-101 has significant synergistic effect with PCI-32765,SAHA and BTZ.