葫芦素B靶向CIP2A介导的C-KIT信号抑制t(8;21)急性髓系白血病细胞增殖并诱导其凋亡  

Cucurbitacin B Targets CIP2A-mediated C-KIT Signal to Inhibit the Proliferation and Induce Apoptosis of t(8;21)Acute Myeloid Leukemia Cells

作　　者：陆俊霏 熊艺琏 欧虹灵 彭鹏 司渊 刘莹 LU Jun-fei;XIONG Yi-lian;OU Hong-ling;PENG Peng;SI Yuan;LIU Ying(School of Basic Medical Sciences,Hu-bei University of Medicine,Shiyan,Hubei 442000,China;Hubei Key Laboratory of Wudang Local Chinese Medicine Research,Hu-bei University of Medicine,Shiyan,Hubei 442000,China)

机构地区：[1]湖北医药学院基础医学院,湖北十堰442000 [2]湖北医药学院武当特色中药研究湖北省重点实验室,湖北十堰442000

出　　处：《湖北医药学院学报》2022年第1期21-27,33,F0002,共9页Journal of Hubei University of Medicine

基　　金：国家自然科学基金(82072928);湖北医药学院研究生科技创新项目(YC2021004);2021年度大学生创新创业训练计划项目(S202110929002)。

摘　　要：目的:本文旨在探讨中药活性成分葫芦素B(cucurbitacin B,CuB)抑制急性髓系白血病(acute myeloid leukemia,AML)的作用及机制。方法:Western blot、QPCR检测蛋白磷酸酶2A的癌性抑制因子(cancerous inhibitor of protein phosphatase 2A,CIP2A)在AML细胞中的表达水平;CCK-8及台盼蓝拒染法检测CuB对AML细胞增殖的抑制作用;DAPI染色和流式细胞术检测CuB对kasumi-1细胞凋亡的影响;Western blot检测CuB对kasumi-1细胞中CIP2A表达及凋亡蛋白表达的影响;QPCR检测CuB对CIP2A基因转录的影响;PP2A活性试剂盒检测CuB对Kasumi-1细胞中PP2A活性的影响;Western blot检测C-KIT及其下游信号分子的变化;CuB与PP2A抑制剂冈田酸(Okadaic acid,OA)共处理细胞检测CuB抑制C-KIT是否与PP2A活性的再激活有关。结果:CIP2A在t(8;21)AML中高表达并与AML预后不良显著相关。CuB可以显著抑制kasumi-1细胞增殖并诱导其发生caspase依赖性的细胞凋亡,该作用机制可能与CIP2A/PP2A/C-KIT信号通路有关。结论:CuB通过抑制AML细胞中CIP2A的表达并重激活PP2A,使C-KIT及其下游信号失活,进而抑制AML细胞增殖并诱导其凋亡。Objective This paper aims to explore the effect and mechanism of cucurbitacin B(CuB),an active component of traditional Chinese medicine,on inhibiting acute myeloid leukemia(AML).Methods Western blot and QPCR were used to detect the expression level of cancerous inhibitor of protein phosphatase2 A(CIP2 A)in AML cells.CCK-8 assay and trypan blue dye exclusion assay were used to detect the effect of CuB on the proliferation of AML cells.DAPI staining and flow cytometry assay were used to detect the effect of CuB on apoptosis of kasumi-1 cells.Western blot was used to detect the effect of CuB on CIP2 A and apoptotic protein in kasumi-1 cells.QPCR was used to detect the effect of CuB on CIP2 A transcription.PP2 A activity kit was uesd to detect the effect of CuB on the activity of PP2 A in kasumi-1 cells.Western blot was used to detect the changes of C-KIT and its downstream signaling molecules.CuB and PP2 A inhibitor Okadaic acid(OA)co-treated cells were used to detect whether CuB inhibits C-KIT due to reactivation of PP2 A activity.Results CIP2 A is highly expressed in t(8;21)AML and is significantly associated with poor prognosis of AML.CuB can significantly inhibit the proliferation of kasumi-1 cells and induce caspase-dependent apoptosis,which may be related to the CIP2 A/PP2 A/C-KIT signaling pathway.Conclusion CuB inhibits the proliferation of AML cells and induces apoptosis by inhibiting the expression of CIP2 A in AML cells and reactivating PP2 A,inactivating C-KIT and its downstream signals.

关 键 词：葫芦素B 急性髓系白血病 CIP2A C-KIT PP2A 

分 类 号：R285[医药卫生—中药学]