HPLC检测白血病细胞系CYP3A5活性方法的建立与应用  

Development and Application of HPLC Assay for Measuring CYP3A5 Activities in Leukemia Cell Lines

作　　者：王婷[1] 陈芳源[1] 韩洁英[1] 钟济华[1] 滕晔[1] 欧阳仁荣[1] 

机构地区：[1]上海第二医科大学仁济医院血液科,上海200001

出　　处：《上海第二医科大学学报》2005年第5期440-442,446,共4页Acta Universitatis Medicinalis Secondae Shanghai

基　　金：国家自然科学基金(30400183)资助项目.

摘　　要：目的建立白血病(AL)细胞系CYP3A5活性检测方法,研究化疗药对其活性的调控。方法HPLC法检测药物干预后AL细胞系CYP3A5活性。结果以1×106细胞、氢化考的松终浓度100μmol/L、孵育24h为AL细胞体外培养条件下氢化考的松6β-羟化活性检测的孵育条件。K562细胞CYP3A5活性显著高于HL-60、NB4和Jurkat细胞。柔红霉素作用24h后K562细胞CYP3A5活性增高,48h后活性显著增高;而NB4与Jurkat细胞在其作用后活性无改变。地塞米松作用24h后Jurkat细胞CYP3A5活性显著增高,48h后活性又显著增高。全反式维甲酸作用24h后NB4细胞CYP3A5活性显著增高,72h后活性又显著增高。结论HPLC检测AL细胞体外培养条件下氢化考的松6β-羟化活性方法的建立可用于研究AL细胞CYP3A5活性。柔红霉素、地塞米松、全反式维甲酸的应用可诱导某些AL细胞株CYP3A5活性。Objective To develope HPLC assay to study CYP3A5 activities regulated by anticancer drugs in leukemia cell lines. Methods CYP3A5 activities of intact leukemia cell lines were detected through HPLC assay. Results 1×10 6 cells incubating with hydrocortisone (HC, final concentration 100 μmol/L) for 24 h were finally determined as the incubation conditions for assessing activities of HC 6β-hydroxylation in intact leukemia cell lines. CYP3A5 activity of K562 cells was statistically significantly higher than that of HL-60, NB4 and Jurkat cells. Treatment with daunorubicin increased CYP3A5 activity after 24 h in K562 cells, and further increased a statistically significantly higher activity after 48 h, while there were no remarkable changes in NB4 and Jurkat cell lines. Dexa- methasone induced a statistically significantly increased CYP3A5 activity after 24 h in Jurkat cells and statistically a even more significantly higher activity after 48 h. All-trans retinoic acid significantly increased CYP3A5 activity after 24 h in NB4 cells statistically, and induced a even more statistically significantly higher activity after 72 h. Conclusion Development of HPLC assay for measuring activities of HC 6β-hydroxylation in intact leukemia cell lines can be used to determine the localized CYP3A5 activities in leukemia cells.Treatment with daunorubicin, dexamethasone and all-trans retinoic acid induced CYP3A5 activities in some leukemia cell lines.

关 键 词：CYP3A5 HPLC检测 白血病细胞系 Jurkat细胞 体外培养条件 K562细胞 全反式维甲酸 氢化考的松 AL细胞 柔红霉素 地塞米松 HPLC法 mol/L HL-60 NB4细胞 24h 检测方法 活性检测 活性增高 72h后 干预后 细胞株 孵育 羟化 β- 

分 类 号：R733.7[医药卫生—肿瘤]