CAJ | 학술논문

目的探讨硼替佐米(Bortezomib)在体外对耐全反式维甲酸(ATRA)的急性早幼粒细胞白血病(APL)细胞株NB_4.R2增殖、凋亡的影响,为用Bortezomib治疗耐ATRA的APL提供新的理论依据.方法体外用不同浓度的Bortezomib处理NB_4-R2细胞株48 h后,用四甲基偶氮唑蓝染色法(MTY)观察细胞增殖活力、Hoechst33342染色法观察细胞凋亡形态、流式细胞术检测细胞凋亡百分率以及Weslern blotting蛋白印迹检测剪切多聚ADP-核糖聚合酶(cleaved PARP)和Caspase-3凋亡蛋白的表达水平.结果体外Bortezomib能明显抑制NB_4-R2细胞增殖,在质量浓度1～5μg/L的范围内,细胞的抑制作用和Bortezomib浓度呈剂量依赖关系,当质量浓度为5μg/L时,细胞增殖抑制率可达74.9%;在质量浓度1～5μg/L的Bonezomih范嗣内,NB_4-R2细胞增殖受抑主要表现为细胞凋亡增加,凋亡百分率可达78.7%,且以晚期凋亡为主;Western blotting证实Bortezomib处理后的NB_4-R2细胞中,cleaved PARP和Caspase-3蛋白表达明显增强.结论体外Bortezomib可通过诱导细胞凋亡来抑制NB_4-R2细胞的增殖.
목적 탐토붕체좌미(Bortezomib)재체외대내전반식유갑산(ATRA)적급성조유립세포백혈병(APL)세포주NB_4.R2증식、조망적영향,위용Bortezomib치료내ATRA적APL제공신적이론의거.방법 체외용불동농도적Bortezomib처리NB_4-R2세포주48 h후,용사갑기우담서람염색법(MTY)관찰세포증식활력、Hoechst33342염색법관찰세포조망형태、류식세포술검측세포조망백분솔이급Weslern blotting단백인적검측전절다취ADP-핵당취합매(cleaved PARP)화Caspase-3조망단백적표체수평.결과 체외Bortezomib능명현억제NB_4-R2세포증식,재질량농도1～5μg/L적범위내,세포적억제작용화Bortezomib농도정제량의뢰관계,당질량농도위5μg/L시,세포증식억제솔가체74.9%;재질량농도1～5μg/L적Bonezomih범사내,NB_4-R2세포증식수억주요표현위세포조망증가,조망백분솔가체78.7%,차이만기조망위주;Western blotting증실Bortezomib처리후적NB_4-R2세포중,cleaved PARP화Caspase-3단백표체명현증강.결론 체외Bortezomib가통과유도세포조망래억제NB_4-R2세포적증식.
Objective To investigate the effect of bortezomib on the proliferation and apoptosis in leukemia cell line NB_4-R2 in vitro and provide some new evidences for the treatment of acute promyelocytic leukemia APL with ATRA-resistant using bortezomib. Methods NB_4-R2 cells were incubated with bortezomib at different does for 48 h. The proliferation capacity was measured by MTT assay, the morphology of cell apoptosis observed with Hoechst33342 staining by fluorescence microscopy and the percentage of apoptosis calculated by flow cytometry. The expression of apoptosis protein of cleaved (poly ADP-ribose polymerase, PARP) and Caspase-3 were determined by Western blotting. Results The proliferation of NB_4-R2 cells were obviously inhibited by bortezomib in vivo and the role of inhibition was a does-dependant manner within the scope of the bortezomib concentration from 1-5 μg /L.The incidence of inhibition was up to 74.9 % at the bortezomib concentration of 5 μg/L. Within this scope of the bortezomib concentration mentioned above, the role of inhibition of proliferation of NB_4-R2 cells mainly showed an increase of the late apoptosis, and the percentage of apoptosis was up to 78.9 %. In the meaning time, the expressions of the apoptotic protein of cleaved PARP and Caspase-3 were up-regulated in NB_4-R2 cells after treated with bortezomib by Western blotting assay. Conclusion Bortezomib can inhibit the proliferation of NB_4-R2 cells in vivo by inducing cell apoptosis.