CAJ | 학술논문

本研究旨在探讨葡萄糖神经酰胺合成酶(glucosylceramide synthase,GCS)抑制剂D,L-threo-1-phenyl-2-de-canoylamino-3-morpholino-1-propanol(PDMP)hydrochloride对K562/A02细胞株柔红霉素(daunorubicin,DNR)耐药的逆转作用及其逆转机制.采用四甲基偶氮唑蓝(MTT)法检测不同浓度PDMP对K562细胞和K562/A02细胞的增殖抑制效应及其对K562/A02细胞DNR耐药逆转作用;用流式细胞术检测PDMP对K562/A02细胞内DNR浓度及细胞凋亡率的影响;半定量RT-PCR和Western blot方法检测K562及K562/A02细胞mdrl、GCS基因的表达以及PDMP对K562/A02细胞两种基因表达的影响.结果表明:DNR对K562和K562/A02细胞的IC50分别为0.23±0.02和7.15±0.24μg/ml,当PDMP≤20 μmol/L时,对K562及K562/A02细胞株无明显生长抑制作用;20 μmol/L和10 μmol/L PDMP均能增加K562/A02细胞对DNR的敏感性,其耐药逆转倍数分别为2.59和1.69;两种浓度PDMP作用于耐药株48小时后能增加细胞内DNR浓度(P＜0.05)和细胞凋亡率(P＜0.01).20 μmol/L PDMP处理K562/A02细胞48小时后可在mRNA和蛋白水平下调GCS和mdr1基因的表达(p＜0.01).结论:PDMP可以增强K562/A02细胞对DNR的敏感性,其机制可能与增加细胞内药物浓度及细胞凋亡率、下调GCS和mdr1基因表达有关.
본연구지재탐토포도당신경선알합성매(glucosylceramide synthase,GCS)억제제D,L-threo-1-phenyl-2-de-canoylamino-3-morpholino-1-propanol(PDMP)hydrochloride대K562/A02세포주유홍매소(daunorubicin,DNR)내약적역전작용급기역전궤제.채용사갑기우담서람(MTT)법검측불동농도PDMP대K562세포화K562/A02세포적증식억제효응급기대K562/A02세포DNR내약역전작용;용류식세포술검측PDMP대K562/A02세포내DNR농도급세포조망솔적영향;반정량RT-PCR화Western blot방법검측K562급K562/A02세포mdrl、GCS기인적표체이급PDMP대K562/A02세포량충기인표체적영향.결과표명:DNR대K562화K562/A02세포적IC50분별위0.23±0.02화7.15±0.24μg/ml,당PDMP≤20 μmol/L시,대K562급K562/A02세포주무명현생장억제작용;20 μmol/L화10 μmol/L PDMP균능증가K562/A02세포대DNR적민감성,기내약역전배수분별위2.59화1.69;량충농도PDMP작용우내약주48소시후능증가세포내DNR농도(P＜0.05)화세포조망솔(P＜0.01).20 μmol/L PDMP처리K562/A02세포48소시후가재mRNA화단백수평하조GCS화mdr1기인적표체(p＜0.01).결론:PDMP가이증강K562/A02세포대DNR적민감성,기궤제가능여증가세포내약물농도급세포조망솔、하조GCS화mdr1기인표체유관.