秦巴硒菇提取物FA-2-b-β介导Wnt信号通路逆转急性T淋巴细胞白血病细胞多药耐药性的机制研究  

Study on the Mechanism of Multi-Drug Resistance of Agaricus Blazei Extract FA-2-b-βMediated Wnt Signaling Pathway to Reverse Acute T Lymphoblastic Leukemia

作　　者：冯文雯 白宇[1] 王东萍 范富艳 孙延庆 FENG Wen-Wen;BAI Yu;WANG Dong-Ping;FAN Fu-Yan;SUN Yan-Qing(The First Clinical Medical College of Gansu University of Traditional Chinese Medicine;Department of Hematology,Gansu Provincial People's Hospital,Lanzhou 730000,Gansu Province,China)

机构地区：[1]甘肃中医药大学第一临床医学院 [2]甘肃省人民医院血液内科,甘肃兰州73000

出　　处：《中国实验血液学杂志》2023年第3期621-627,共7页Journal of Experimental Hematology

基　　金：甘肃省自然科学基金(项目编号:20JR10RA376、21JR11RA196)。

摘　　要：目的:探讨秦巴硒菇提取物FA-2-b-β对急性T淋巴细胞白血病细胞的逆转耐药作用机制。方法:采用CCK-8法检测细胞增殖;流式细胞术检测细胞凋亡、线粒体膜电位、细胞周期以及胞内罗丹明蓄积情况;qPCR和Western blot检测凋亡相关基因与蛋白表达;免疫荧光观察膜表面蛋白MDR1表达。结果:不同浓度秦巴硒菇提取物FA-2-b-β能够显著抑制CCRF-CEM及CEM/C1细胞增殖(P<0.05),促进其凋亡,并调控CCRF-CEM细胞阻滞于S期、CEM/C1细胞阻滞于G_(0)/G_(1)期。Western blot及qPCR检测结果显示,FA-2-b-β可抑制ABCB1、ABCG_(2)、CTNNB、MYC、BCL-2表达,并上调BAX表达。另外,FA-2-b-β可逆转CEM/C1耐药株的耐药特性,使得其线粒体膜电位下降,细胞内罗丹明蓄积明显增多,膜表面蛋白MDR1表达减弱。加入ICG_(0)01后,该过程进一步加强。结论:秦巴硒菇提取物FA-2-b-β通过抑制细胞增殖促进凋亡、调控细胞周期、减少线粒体供能及下调MDR1蛋白表达等方式逆转CEM/C1细胞的耐药性,其机制可能通过调控Wnt信号通路实现。Objective:To investigate the mechanism of drug reversing resistance of Agaricus blazei extract FA-2-b-βon T cell acute lymphoblastic leukemia(T-ALL)cell lines.Methods:Cell proliferation was detected by CCK-8 assay;the apoptosis,cell cycle mitochondrial membrane potential,and intracellular rhodamine accumulation were detected by flow cytometry,and apoptosis-related gene and protein expression were detected by qPCR and Western blot;the membrane surface protein MDR1 was observed by immunofluorescence microscopy.Results:Different concentrations of FA-2-b-βsignificantly inhibited proliferation and induced apoptosis of CCRF-CEM and CEM/C1(P<0.05),and CCRF-CEM cell cycle were arrested at S phase,and CEM/C1 cells were arrested at G_(0/)G_(1) phase.Western blot and qPCR results show that FA-2-b-βinhibited ABCB1、ABCG2、CTNNB、MYC and BCL-2 expression,but upregulated Bax expression.In addition,FA-2-b-βreversed the resistance characteristics of CEM/C1 drug-resistance cells,which decreased mitochondrial membrane potential,and significantly increased the intracellular rhodamine accumulation,and weakening of the expression of the membrane surface protein MDR1.With the Wnt/β-catenin inhibitor(ICG001),the process was further intensified.Conclusion:Agaricus Blazei Extract FA-2-b-βinhibits cell proliferation,promotes apoptosis,regulates the cell cycle,reduces mitochondrial energy supply,and down-regulate MDR1 expression to reverse the resistance of CEM/C1,which all suggest it is through regulating the Wnt signaling pathway in T-ALL.

关 键 词：秦巴硒菇 急性T淋巴细胞白血病 多药耐药 WNT信号通路 

分 类 号：R733.71[医药卫生—肿瘤]