CAJ | 학술논문

目的:观察环孢素A(Cyclosporin A,CsA)处理后急性单核细胞白血病细胞株THP-1多药耐药相关蛋白-1(Multidrug resistance associated protein-1,MRP1)表达量和活性的变化,探讨CsA逆转MRP1的相关机制及THP-1细胞对柔红霉素(Daunorubicin.DNR)的增敏作用.方法:免疫细胞化学法和Western blot法检测CsA处理后THP-1细胞MRP1表达量的改变,噻唑蓝(MTT)法、荧光分光光度法、共聚焦激光扫描显微镜(Confocal laser scanning microscopy,CLSM)法分别检测CsA处理后50%THP-1细胞生长受抑的DNR浓度(IC50)、THP-1细胞内DNR含量及分布.结果:CsA处理组较对照组MRP1表达量定性、定量检测均增加(P＜0.05);IC50明显下降(0.558 4±0.0670与1.4997±0.2338,P＜0.01),细胞内DNR含量明显增加(126.500 0±14.781 5与20.9000 4-2.497 5,P＜0.01),分布更加均匀.结论:CsA增加MRP1的表达量,但降低其活性,提示CsA通过降低MRP1活性而非减少其表达量实现对MRP1的逆转作用和对THP-1细胞的增敏作用
목적:관찰배포소A(Cyclosporin A,CsA)처리후급성단핵세포백혈병세포주THP-1다약내약상관단백-1(Multidrug resistance associated protein-1,MRP1)표체량화활성적변화,탐토CsA역전MRP1적상관궤제급THP-1세포대유홍매소(Daunorubicin.DNR)적증민작용.방법:면역세포화학법화Western blot법검측CsA처리후THP-1세포MRP1표체량적개변,새서람(MTT)법、형광분광광도법、공취초격광소묘현미경(Confocal laser scanning microscopy,CLSM)법분별검측CsA처리후50%THP-1세포생장수억적DNR농도(IC50)、THP-1세포내DNR함량급분포.결과:CsA처리조교대조조MRP1표체량정성、정량검측균증가(P＜0.05);IC50명현하강(0.558 4±0.0670여1.4997±0.2338,P＜0.01),세포내DNR함량명현증가(126.500 0±14.781 5여20.9000 4-2.497 5,P＜0.01),분포경가균균.결론:CsA증가MRP1적표체량,단강저기활성,제시CsA통과강저MRP1활성이비감소기표체량실현대MRP1적역전작용화대THP-1세포적증민작용
Objective: To observe the changes of expression and activity of multidrug resistance associated protein-1 ( MRP1) in acute monocytic leukemia THP-1 cells treated by cyclosporin A(CsA), and to explore the mechanism that the function of MRP1 can be reversed by CsA making the THP-1 cells sensiting to chemotherapy. Methods: Expression of MRP1 in THP-1 cells treated by CsA is detected by Immunocytochemistry and Western blot in sequence.50% inhibiting concentration (IC50) of daunorubicin(DNR) to THP-1 cells, content and distribution of DNR in THP-1 cells treated by CsA are examined by MTT method, Spectrofluorometer method and Confocal laser scanning microscopy method,respectively. Results:Compaed with Control Group, in Group CsA the expression of MRP1 increased both in quality while quantity(P＜0.05) and IC50, decreased obviously(1.499 7 ± 0.233 8 vs 0.558 4 ± 0.067 0,P＜0.01);content of DNR increased obviously(20.900 0± 2.497 5 vs 126.500 0± 14.781 5,P＜0.01),and DNR distributed more evenly. Conclusion:CsA can increase the expression of MRP1 and decreased its activity, showing that the resistance of MRP1 is reduced by decreasing the activity of MRP1 ,not the amount of expression, as well as increasing chemotherapy sensitivity of THP-1 cells.