CAJ | 학술논문

目的：研究五味子乙素（schisandrin B，SchB）对人骨肉瘤细胞阿霉素耐药株U-2 OS／ADR多药耐药的逆转效果及逆转机制。方法采用浓度梯度递增法构建U-2 OS阿霉素耐药株U-2 OS／ADR；使用MTT法测定Sch B对于U-2 OS多药耐药逆转的影响，实时荧光定量PCR（ Q-PCR）检测SchB对MDR1基因转录的影响，流式细胞术检测细胞膜表面MDR1蛋白的表达，流式细胞术检测五味子乙素对罗丹明123外排和蓄积的影响，Western Blotting检测五味子乙素对PI3K／AKT通路的影响。结果五味子乙素能逆转U-2 OS／ADR细胞的多药耐药，并且能抑制MDR1基因的转录，降低膜表面MDR1蛋白的表达，增加细胞内罗丹明123的蓄积、减少外排，抑制U-2 OS／ADR细胞中PI3K／AKT通路的激活。结论五味子乙素具有强大的逆转人骨肉瘤细胞U-2 OS多药耐药的效果，其机制和下调耐药株的MDR1基因和蛋白水平，抑制PI3K／AKT通路激活有关。
목적：연구오미자을소（schisandrin B，SchB）대인골육류세포아매소내약주U-2 OS／ADR다약내약적역전효과급역전궤제。방법채용농도제도체증법구건U-2 OS아매소내약주U-2 OS／ADR；사용MTT법측정Sch B대우U-2 OS다약내약역전적영향，실시형광정량PCR（ Q-PCR）검측SchB대MDR1기인전록적영향，류식세포술검측세포막표면MDR1단백적표체，류식세포술검측오미자을소대라단명123외배화축적적영향，Western Blotting검측오미자을소대PI3K／AKT통로적영향。결과오미자을소능역전U-2 OS／ADR세포적다약내약，병차능억제MDR1기인적전록，강저막표면MDR1단백적표체，증가세포내라단명123적축적、감소외배，억제U-2 OS／ADR세포중PI3K／AKT통로적격활。결론오미자을소구유강대적역전인골육류세포U-2 OS다약내약적효과，기궤제화하조내약주적MDR1기인화단백수평，억제PI3K／AKT통로격활유관。
Objective To study the reversal effect of schisandrin B on doxorubicin induced drug-fast human osteosarcoma cell line U-2 OS/ADR.Methods U-2 OS/ADR was established by increasing the concentration gradient of doxorubicin in a stepwise manner ;deter-mine the effects of SchB on reversal of multidrug resistance by MTT , test the effect of SchB on the Gene transcription of MDR 1 by quantitative real-time PCR, test the expression of MDR1 on the cell membrane as well as the influence of SchB on the excretion and ac-cumulation of Luo Danming123 by the method of flow cytometry , test schisandrin B'effect on PI3K/AKT signal pathway by western blotting.Results Schisandrin B showed potent reversal effect on multidrug resistance (MDR) of U-2 OS/ADR cell and has the func-tion of inhibiting the transcription of MDR1 gene, reducing the expression of MDR1 on cell membrane, increasing the accumulation and decreasing the excretion of Luo Danming 123 inside of the cell, as well as inhibiting the activation of PI 3K/AKT signal pathway in U-2 OS/ADR.Conclusions Schisandrin B could reverse the MDR of U-2 OS/ADR by inhibiting the MDR1 and the PI3K/AKT signaling.