榄香烯注射液对吉非替尼在PC-9/GR细胞内转运动力学影响

Effects of Elemene Injection on the kinetics of intracellular transport of Gefitinib in PC-9/GR cells

目的研究榄香烯注射液对吉非替尼(GEF)在PC-9/GR细胞内转运动力学影响,探讨该药在逆转肿瘤多药耐药方面的作用。方法活体生物发光技术实时监测PC-9/GR^(Fluc)细胞内ABC耐药蛋白底物D-荧光素钾动力学行为。采用MTT法测定PC-9/GR细胞的耐药性,榄香烯、GEF单用药及联合用药时的细胞毒性,Compusyn软件分析两药联合是否具有协同作用。运用HPLC法,测定联合用药组及GEF单用药组药物在PC-9/GR细胞中的摄取量。结果榄香烯能显著增加细胞内D-荧光素钾的荧光响应强度。吉非替尼对PC-9及PC-9/GR细胞的IC50值分别为0.01、1.50μg/m L,耐药倍数为150倍。所有联合药物组的联合用药指数(combination index,CI)值都小于1,表明两药联合具有较好的协同作用。细胞摄取结果表明,配伍榄香烯(ELE)后,GEF在PC-9/GR细胞摄取明显增加(P<0.05)。结论榄香烯增加D-荧光素钾荧光响应强度,促进细胞对GEF摄取,说明在一定程度上能抑制耐药蛋白的外排功能从而逆转耐药。

AIM To study the effects of Elemene Injection（ ELE） on the kinetics of intracellular transport of Gefitinib（ GEF） in PC-9/GR cells and to probe the role of ELE in reversing oncological multidrug resistance.METHODS The intracellular pharmacokinetic behavior of D-luciferin potassium salt,a substrate of an ATP-binding cassette（ ABC） protein,was investigated in PC-9/GR^（Fluc） cells using real-time bioluminescence imaging. The resistance of PC-9/GR cells to GEF was determined by MTT assay. Compusyn software was used to analyze the synergistic effect of GEF and ELE,and HPLC to detect the uptake of GEF in PC-9/GR cells. RESULTS The respective GEF IC50 values of 0. 01 μg/mL in PC-9 cells and 1. 50 μg/m L in PC-9/GR cells revealed the 150 times drug resistance of PC-9/GR to PC-9 cells. The significantly enhanced intracellular fluorescence intensity of D-fluorescein potassium salt by the intervention of ELE also indicated remarkable GEF uptake increase in PC-9/GR cell line（ P〈0. 05） due to the synergistic result. CONCLUSION Partly as the mechanism in reversing oncological multidrug resistance,ELE,a booster for the fluorescence intensity of D-luciferin potassium salt,promotes cellular uptake of GEF by inhibiting efflux function of ABC proteins.