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

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Citation abstract

OBJECTIVE: To investigate the mechanism of drug reversing resistance of Agaricus blazei extract FA-2-b-β on 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-β significantly 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-β inhibited ABCB1、ABCG2、CTNNB、MYC and BCL-2 expression, but upregulated Bax expression. In addition, FA-2-b-β reversed 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-β inhibits 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.

题目: 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.

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、ABCG2、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ICG001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.