秦巴硒菇提取物联合多柔比星抗白血病K562/ADR细胞多药耐药的机制研究  

Agaricus blazei Murrill Extract Combined with Adriamycin Inhibits Multidrug Resistance of K562/ADR Cells

作　　者：王东萍 葛万文 董莉[3] 孙延庆 WANG Dongping;GE Wanwen;DONG Li;SUN Yanqing(School of Traditional Chinese and Western Medicine,Gansu University of Chinese Medicine,Gansu Provincial Hospital,Lanzhou 730000;Cuiying Biomedical Research Center,Lanzhou University Second Hospital,Lanzhou 730030;Gansu Provincial Hospital,Lanzhou 730000)

机构地区：[1]甘肃中医药大学中西医结合学院,甘肃省人民医院,兰州730000 [2]兰州大学第二医院萃英生物医学研究中心,兰州730030 [3]甘肃省人民医院,兰州730000

出　　处：《中药药理与临床》2024年第2期67-72,共6页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金(编号:81560670);甘肃省自然科学基金(编号:20JR10RA376、21JR11RA196);兰州市科技发展指导性计划项目(编号:2020-ZD-56);兰州大学第二医院“萃英科技创新”计划项目(编号:CY-2019-QN15);甘肃省高等学校创新基金项目(编号:2020B-040)。

摘　　要：目的:探讨秦巴硒菇提取物FA-2-b-β联合多柔比星(Adriamycin, ADR)抗白血病K562/ADR细胞多药耐药的协同抗肿瘤机制。方法:通过转录组测序分析筛选白血病敏感细胞K562和耐药细胞K562/ADR中的差异表达基因,分析可能的信号通路;CCK-8法检测K562/ADR细胞的耐药表型及FA-2-b-β对多柔比星的增敏作用;流式细胞术检测不同药物干预后各组细胞的凋亡比例、细胞内多柔比星的荧光强度变化;蛋白免疫印迹法(Western blot)检测耐药蛋白、凋亡相关蛋白以及PI3K/AKT/mTOR信号通路相关蛋白的表达。结果:与敏感细胞K562相比,K562/ADR细胞的耐药性明显增加,耐药倍数为26.88倍。试验结果表明,与空白对照组相比,FA-2-b-β协同多柔比星对K562/ADR细胞的增殖有明显抑制作用,可显著诱导K562/ADR细胞凋亡,并提高细胞内多柔比星的荧光强度;上调促凋亡蛋白BAD的表达,明显下调耐药蛋白P-gp、抗凋亡蛋白BCL-XL以及p-PI3K、p-AKT和p-mTOR蛋白的表达(P<0.05)。结论:秦巴硒菇提取物FA-2-b-β联合多柔比星有协同抗肿瘤作用,可能是通过促进K562/ADR细胞凋亡、减少细胞内化疗药物外排并抑制耐药蛋白而发挥作用,其作用机制可能与调控PI3K/AKT/mTOR通路有关。Objective: To investigate the mechanism of Agaricus blazei Murrill extract FA-2-b-βcombined with adriamycin (ADR) in inhibiting the multidrug resistance of K562/ADR cells.Methods: Transcriptome sequencing was employed to identify the differentially expressed genesbetween the sensitive cell line K562 and the drug-resistant cell line K562/ADR and reveal thepossible signaling pathways. The cell-counting kit 8 (CCK-8) was used to examine the drugresistance phenotype of K562/ADR cells and FA-2-b-β sensibilization on ADR. Flow cytometrywas employed to detect the apoptosis of K562/ADR cells treated with different drugs and themean fluorescence intensity (MFI) of ADR. Western blotting was employed to determine theexpression levels of the proteins involved in drug resistance, apoptosis, and phosphatidylinositol3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signalingpathway. Results: Compared with K562 cells, the resistance of K562/ADR cells increased by26.88 times. Compared with the blank control group, FA-2-b-β in combination with ADRincreased the inhibitory effect on cell growth, induced cell apoptosis, and increased the MFI ofADR in K562/ADR cells. Moreover, the combination up-regulated the expression of the apoptosisprotein BAD and down-regulated the protein levels of P-gp, the anti-apoptotic protein BCL-XL,p-PI3K, p-Akt, and p-mTOR (P<0.05). Conclusion: A. blazei extract FA-2-b-β combined withADR exerts synergistic anti-tumor effects on K562/ADR cells by inducing cell apoptosis,down-regulating the expression of resistance proteins, and inhibiting drug efflux, which may beassociated with the down-regulation of the PI3K/Akt/mTOR signaling pathway.

关 键 词：秦巴硒菇提取物 多柔比星 白血病 耐药 磷脂酰肌醇-3激酶/蛋白激酶/雷帕雷素 

分 类 号：R285[医药卫生—中药学]