全反式维甲酸与阿糖胞苷或柔红霉素不同联用方式对体外HL-60氧张力及凋亡相关通路的影响  

Different ways of combination of all-trans retinoic acid with cytarabine or daunorubicin lead to variable changes of HL-60's oxygen tension and signal pathways in apoptosis regulation in vitro

作　　者：陈敏玲 黄诗颖 张顺国 吴娟 

机构地区：[1]上海交通大学医学院附属上海儿童医学中心药剂科,上海200127

出　　处：《中国临床药学杂志》2013年第4期201-206,共6页Chinese Journal of Clinical Pharmacy

基　　金：上海交通大学医学院药学科研基金(编号JYY0912)

摘　　要：目的研究全反式维甲酸(ATRA)以不同方式联用阿糖胞苷(Ara-C)或柔红霉素(DNR),作用于白血病细胞株HL-60致细胞内氧化还原物质及凋亡相关通路蛋白的变化。方法体外培养HL-60细胞株,细胞悬液以密度5×10~5·mL^(-1)接种于24孔板中。将细胞分为7组,A组为正常对照组;B组将ATRA与Ara-C同时联合作用24 h;C组预先用ATRA作用24 h,然后改用Ara-C继续作用24h;D组预先用Ara-C作用24h,然后改用ATRA继续作用24h;E组将ATRA与DNR同时联合作用24h;F组预先用ATRA作用24h,然后改用DINR继续作用24h;G组预先用DNR作用24h,然后改用ATRA继续作用24h。MTT法测定细胞活力,根据试剂盒说明书测定细胞内总SOD酶活力和GSH水平,Westem印迹检测唯BH3域蛋白Bim表达量及MAPK信号通路磷酸化水平。结果细胞抑制率为序贯给药组(C、D、G、F组)高于同时给药组(B、E组),在序贯给药组中为先化疗药后ATRA组(D、G组)高于先ATRA后化疗药(C、F组)。与A组相比,各组SOD酶活力均明显降低,其中C、F组降低最多。D、G组细胞内GSH水平明显降低,而B、C、E、F组GSH水平明显升高。唯BH3域蛋白Bim相对表达水平随细胞抑制率升高而增多。B、C、E、F组JNK磷酸化水平降低,D、G组升高;B、C、E组p38磷酸化增多;各给药组ERK磷酸化情况无显著变化。结论不同的药物联用方式能通过细胞内GSH水平、凋亡相关蛋白以及MAPK信号通路来影响HL-60的凋亡水平。AIM To investigate variable change of oxygen tension and signal pathways in apoptosis regulation by different ways of combination of all-trans retinoic acid （ATRA） with cytarabine （Ara-C） or daunorubicin （DNR） to HL- 60 cell line in vitro. METHODS HL-60 cell line was cultured in vitro, cell suspension was divided into different 7 groups： group A - G, which respectively were： control （group A） ; Ara-C and ATRA simultaneously were applied in cells for 24 h （group B） ; ATRA and Ara-C subsequently were administrated respectively for 24 h including ATRA was applied first for 24 h, then followed by Ara-C for 24 h （group C） ,and Am-C was applied for 24 h,then followed by A- TRA for 24 h （group D）; DNR and ATRA simultaneously were applied for 24 h （group E）; DNR and ATRA subse- quently were administrated respectively for 24 h （group F, group G）. The cell viability was evaluated by MTF assay. The total supemxide dismutase （SOD） enzyme activity and glutathione （GSH） level were observed to determine the effects of different administration of combination on oxygen tension. The BH3-only protein Bim, the pm-apoptosis protein, and the MAPK signal pathway were also determined by Western Blotting. RESULTS Group D and G showed the highest in- hibitory ratio and there was the lowest inhibitory ratio in group B and E. Compared with group A, SOD activity was sig- nificantly decreased in group B - G, with the lowest level in group C and F. The level of GSH was greatly increased in group B, C, E and F, but decreased in group D and G. There was a positive correlation between the expression of pro- apoptosis protein Bim and the inhibitory ratio. Phosphorylation of JNK in group B, C, E and F was inhibited, but the promoted phosphorylation was found in group D and G. p38 phosphorylation was remarkable increased in group B, C and E. However, no significant difference in ERK activation was found within all the groups when compared to group A. CONCLUSION Different combination of drugs can affect

关 键 词：全反式维甲酸 阿糖胞苷 柔红霉素 HL-60 氧张力 唯BH3域蛋白 MAPK通路 

分 类 号：R965[医药卫生—药理学]