摘要
[目的]探讨TopoⅡβ在苦参碱诱导维甲酸耐药急性早幼粒细胞白血病(acute promye locyticleukemia,APL)细胞分化的作用。[方法]以全反式维甲酸(all-trans retinoic acid,ATRA)敏感的APL细胞系NB4及其ATRA耐药细胞株NB4-R1作为研究对象,选取对数生长期细胞,分为空白细胞对照组、ATRA组、苦参碱组、ATRA+苦参碱组。培养3d后,研究各组细胞形态的变化;用NBT还原实验分析细胞分化能力的变化;流式细胞术检测细胞表面分化抗原CD11b的表达变化;免疫组化法检测拓扑异构酶TopoⅡβ亚型的表达变化。[结果]与ATRA组相比,0.1mmo.lL-1苦参碱可协同1μmo.lL-1维甲酸诱导NB4-R1细胞分化,使NBT还原阳性率明显增高(12%VS 41.33%),并能诱导NB4-R1细胞表面分化抗原CD11b的表达百分率升高(32.776±6.610 VS 46.888±7.847)%;而对NB4细胞则无明显变化。单用苦参碱对NB4及NB4-R1细胞内TopoⅡβ均无明显作用;维甲酸联合苦参碱能降低耐药细胞株NB4-R1细胞内TopoⅡβ含量(P<0.05)。[结论]苦参碱体外能协同ATRA使NB4-R1细胞分化成熟,其作用机制可能与调控TopoⅡβ的表达有关。
[Objective] To explore the effect of Topo Ⅱ β on the differentiation of retinoic acid resistant acute promyelocytic leukemia induced by matrine.[Methods] ATRA sensitive strain of APL（NB4） and resistant strain（NB4-R1） were used in this study.The logarithmic growth phase cells of each cell line were divided into four groups,i.e.,the control group,the ATRA group,the matrine（MAT） group,the ATRA ＋ MAT group.Each group was cultured for 72 hours.Cell morphologic changes were observed under optical microscope.The influence of MAT combines with ATRA on the differentiation of the two cell strains was observed by nitro blue tetrazolium chioride（NBT） test.Cell surface differentiation antigens CD11b expression was measured by flow cytometry.The protein expression of topoisomerase Ⅱ β isoform was detected by immunohistochemical assay.[Results] Compared with the ATRA group,0.1mmol /L matrine collaborative 1μmol·L-1 retinoic acid could induce NB4-R1 cells to terminal cell differentiation,significantly increase the NBT-positive rate（12% VS 41.33%）,and could increase CD11b expression percentage（23.455 ± 5.933 VS 46.888 ± 7.847）%.But to NB4 cells,there was no obvious change between the ATRA group and the ATRA ＋ matrine group.Using matrine on NB4 and NB4-R1,there was no significant effect for the Topo Ⅱ β protein expression.Retinoic acid combined with matrine could reduce intracellular Topo Ⅱ β content of NB4-R1 cell line.[Conclusion] Matrine is synergistic alltrans retinoic acid NB4-R1 cells differentiation,and its mechanism may be related to Topo Ⅱ β expression and regulation.
出处
《浙江中医药大学学报》
CAS
2012年第12期1255-1260,共6页
Journal of Zhejiang Chinese Medical University
基金
浙江省自然科学基金项目(Y2101416)
浙江省教育厅科研计划项目(Y201223638)~~
关键词
苦参碱
APL
维甲酸耐药
TopoⅡβ
分化
Matrine
APL
retinoic acid resistant
Topoisomerase Ⅱ β
differentiation
