摘要
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。
Objective To study the reversal effect of schisandrin B on doxorubicin induced drug-fast human osteosarcoma cell line U-2 OS/ADR. Methods U-20S/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 MDRI 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 accumulation of Luo Danming123 by the method of flow cytometry , test schisandrin B＇effect on PI3K/AKT signal pathway by western blotting. Results Sehisandrin B showed potent reversal effect on multidrug resistance （MDR） of U-20S/ADR cell and has the function of inhibiting the transcription of MDR1 gene, reducing the expression of MDR1 on cell membrane, increasing the accumulation and decreasing the excretion of Luo Danming123 inside of the cell, as well as inhibiting the activation of PI3K/AKT signal pathway in U-2 OS/ADR. Conclusions Schisandrin B could reverse the MDR of U-20S/ADR by inhibiting the MDR1 and the PI3K/AKT signaling.
出处
《安徽医药》
CAS
2014年第9期1642-1645,共4页
Anhui Medical and Pharmaceutical Journal
关键词
五味子乙素
多药耐药
多药耐药蛋白1
schisandrin B
muhidrug resistance
muhidrug resistance protein 1
