知母皂苷AⅢ对急性B淋巴细胞白血病细胞的抑制作用及其号转导机制  

Inhibitory effect of Timosaponin A-Ⅲ on B-acute lymphoblastic leukemia cells and its signal transduction mechanism

作　　者：张艳[1] 刘洪涛 王海桥[3] ZHANG Yan;LIU Hongtao;WANG Haiqiao(Department of Pediatrics,Huantai County People's Hospital,Zibo 256400,China;Department of Radiology,Huantai County Psychiatric Hospital,Zibo 256400,China;Department of Traditional Chinese Medicine,Renji Hospital,School of Medicine,Shanghai Jiaotong University,Shanghai 201112,China)

机构地区：[1]桓台县人民医院儿科,淄博256400 [2]桓台县精神病医院放射科,淄博256400 [3]上海交通大学医学院附属仁济医院中医科,上海201112

出　　处：《免疫学杂志》2020年第10期890-896,共7页Immunological Journal

基　　金：上海市卫健委中医药传承和科技创新项目(ZYCC2019025)。

摘　　要：目的研究知母皂苷AⅢ(TA-Ⅲ)对急性B淋巴细胞白血病(B-ALL)细胞的影响和潜在的作用机制。方法采用不同浓度的TA-Ⅲ对B-ALL细胞进行处理,MTT法检测细胞活力,流式细胞术检测细胞凋亡。Western blot方法检测线粒体途径凋亡信号通路相关蛋白caspase-8、BID、Bax、Bcl-2、细胞色素c、caspase-9、caspase-3、PARP等表达水平和裂解激活水平,并检测PI3K/AKT信号通路相关蛋白AKT,GSK3、FOXO1的磷酸化水平。结果 TA-Ⅲ以剂量依赖方式显著抑制B-ALL细胞活力,促进其凋亡;TA-Ⅲ能剂量依赖性的使caspase-8,caspase-9,caspase-3,PARP裂解激活,上调Bax/Bcl-2比值,并升高细胞质中的细胞色素c表达水平。而caspase抑制剂能显著的抑制这种作用。此外,不同浓度的TA-Ⅲ能提高AKT,GSK3、FOXO1的磷酸化水平。结论 TA-Ⅲ能够抑制B-ALL细胞活力,可能是通过线粒体途径激活caspase级联反应而诱导B-ALL细胞凋亡来完成。这些作用可能与其抑制PI3k/AKT信号通路的激活有关。The aim of this study is to investigate the effect of Timosaponin A-Ⅲ(TA-Ⅲ) on B-acute lymphoblastic leukemia(B-ALL) cells and its potential mechanism. B-ALL cells was treated with different concentrations of TA-Ⅲ, and then the cell viability and apoptosis were detected by MTT and flow cytometry,respectively. Western blot was used to detect the expression levels of caspase-8, BID, bax, Bcl-2, cytochrome C,caspase-9, caspase-3 and PARP, as well as the phosphorylation levels of PI3 K/Akt signal pathway related proteins Akt, GSK3 and FOXO1. Data showed that TA-Ⅲ significantly inhibited the viability of B-ALL cells and induced their apoptosis in a dose-dependent manner. Moreover, TA-Ⅲ could dose-dependently activate the cleavage of caspase-8, caspase-9, caspase-3 and PARP, up-regulate the ratio of Bax/Bcl2, and increase the expression level of cytochrome C in the cytoplasm. However, caspases inhibitor can significantly inhibit these effects mentioned above.In addition, different concentrations of TA-Ⅲ could up-regulate the phosphorylation levels of Akt, GSK3 and FOXO1. Therefore, we speculate that TA-Ⅲ may induce apoptosis of B-ALL cells by activating caspase cascade through mitochondrial pathway. These effects may be related to the inhibition of activation of PI3 K/Akt signaling pathway.

关 键 词：急性B淋巴细胞白血病 知母皂苷AⅢ 细胞凋亡 信号通路 

分 类 号：R733.7[医药卫生—肿瘤]