[1]赵小强,吴雅莉,仝佳音,等.鱼藤素对急性髓系白血病KG-1a细胞增殖和凋亡的影响及机制[J].新乡医学院学报,2021,38(12):1121-1127.[doi:10.7683/xxyxyxb.2021.12.003]
 ZHAO Xiaoqiang,WU Yali,TONG Jiayin,et al.Effect of deguelin on the proliferation and apoptosis of acute myeloid leukemia KG-1a cells and its mechanism[J].Journal of Xinxiang Medical University,2021,38(12):1121-1127.[doi:10.7683/xxyxyxb.2021.12.003]
点击复制

鱼藤素对急性髓系白血病KG-1a细胞增殖和凋亡的影响及机制
分享到:

《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
38
期数:
2021年12
页码:
1121-1127
栏目:
基础研究
出版日期:
2021-12-05

文章信息/Info

Title:
Effect of deguelin on the proliferation and apoptosis of acute myeloid leukemia KG-1a cells and its mechanism
作者:
赵小强吴雅莉仝佳音席晓平程英英杨海平
(河南科技大学第一附属医院/河南科技大学临床医学院血液内科,河南 洛阳 471000)
Author(s):
ZHAO XiaoqiangWU YaliTONG JiayinXI XiaopingCHENG YingyingYANG Haiping
(Department of Hematology,the First Affiliated Hospital of Henan University of Science and Technology/Clinical School of Henan University of Science and Technology,Luoyang 471000,Henan Province,China)
关键词:
急性髓系白血病鱼藤素增殖凋亡
Keywords:
acute myeloid leukemiadeguelinproliferationapoptosis
分类号:
R733.7
DOI:
10.7683/xxyxyxb.2021.12.003
文献标志码:
A
摘要:
目的 探讨鱼藤素对急性髓系白血病(AML)KG-1a细胞增殖、凋亡的影响及机制。方法 将对数生长期KG-1a细胞随机分为0.00 nmol·L-1鱼藤素组、10.00 nmol·L-1鱼藤素组、20.00 nmol·L-1鱼藤素组、40.00 nmol·L-1鱼藤素组、80.00 nmol·L-1鱼藤素组,分别用含有0.00、10.00、20.00、40.00、80.00 nmol·L-1鱼藤素的培养基培养。鱼藤素干预48 h后,采用倒置显微镜观察5组细胞的形态学变化。采用二甲基噻唑(MTT)实验检测5组细胞增殖能力。另取对数生长期KG-1a细胞随机分为空白组、鱼藤素组、氯化锂(LiCl)组、鱼藤素+LiCl组,空白组细胞用正常培养基培养,鱼藤素组细胞用含80.00 nmol·L-1鱼藤素的培养基干预72 h,LiCl组细胞用含30 mmol·L-1 LiCl的培养基干预72 h,鱼藤素+LiCl组细胞用含80.00 nmol·L-1鱼藤素和30 mmol·L-1 LiCl的培养基干预72 h。采用流式细胞术检测4组细胞凋亡情况及细胞周期分布。采用实时荧光定量聚合酶链反应(qRT-PCR)法检测4组细胞S期激酶相关蛋白2(SKP2)、β-链接素(β-catenin)、聚腺苷酸二磷酸核糖转移酶(PARP) mRNA相对表达量。采用Western blot法检测4组细胞中SKP2、β-catenin、PARP蛋白相对表达量。结果 鱼藤素干预48 h时,与0.00 nmol·L-1鱼藤素组比较,10.00 nmol·L-1鱼藤素组、20.00 nmol·L-1鱼藤素组、40.00 nmol·L-1鱼藤素组、80.00 nmol·L-1鱼藤素组细胞间隙增大,细胞数量显著减少,形态不规则,细胞核固缩、碎片增加,且随着鱼藤素药物干预浓度的增加,细胞数量逐渐减少,形态变化逐渐明显。鱼藤素干预24、48、72 h时,10.00 nmol·L-1鱼藤素组、20.00 nmol·L-1鱼藤素组、40.00 nmol·L-1鱼藤素组、80.00 nmol·L-1鱼藤素组细胞增殖能力显著低于0.00 nmol·L-1鱼藤素组(P<0.05),20.00 nmol·L-1鱼藤素组、40.00 nmol·L-1鱼藤素组、80.00 nmol·L-1鱼藤素组细胞增殖能力显著低于10.00 nmol·L-1鱼藤素组(P<0.05),40.00 nmol·L-1鱼藤素组、80.00 nmol·L-1鱼藤素组细胞增殖能力显著低于20.00 nmol·L-1鱼藤素组(P<0.05),80.00 nmol·L-1鱼藤素组细胞增殖能力显著低于40.00 nmol·L-1鱼藤素组(P<0.05);鱼藤素干预24、48、72 h时,5组细胞的增殖能力均随培养时间延长显著降低,组内各时间点之间两两比较差异有统计学意义(P<0.05)。空白组、鱼藤素组、LiCl组、鱼藤素+LiCl组细胞凋亡率分别为(5.37±0.61)%、(34.89±4.57)%、(2.54±0.36)%、(21.10±4.69)%;空白组、LiCl组、鱼藤素+LiCl组细胞凋亡率显著低于鱼藤素组(P<0.05);空白组、LiCl组细胞凋亡率均显著低于鱼藤素+LiCl组;LiCl组细胞凋亡率均显著低于空白组(P<0.05)。空白组、鱼藤素+LiCl组、鱼藤素组的G0/G1 细胞占比显著低于LiCl组,G2/M细胞占比显著高于LiCl组(P<0.05);鱼藤素+LiCl组、鱼藤素组的G0/G1 细胞占比显著低于空白组,G2/M细胞占比显著高于空白组(P<0.05);鱼藤素组的G0/G1 细胞占比显著低于鱼藤素+LiCl组,G2/M细胞占比显著高于鱼藤素+LiCl组(P<0.05);4组S期细胞百分比比较差异无统计学意义(F=0.696,P>0.05)。空白组、鱼藤素+LiCl组、鱼藤素组细胞SKP2、β-catenin、PARP mRNA及蛋白相对表达量显著低于LiCl组(P<0.05),鱼藤素+LiCl组、鱼藤素组细胞SKP2、β-catenin、PARP mRNA及蛋白相对表达量显著低于空白组(P<0.05),鱼藤素组细胞SKP2、β-catenin、PARP mRNA及蛋白相对表达量显著低于鱼藤素+LiCl组(P<0.05)。结论 鱼藤素对AML KG-1a细胞具有增殖抑制、细胞周期阻滞及凋亡促进作用,其作用机制可能与抑制SKP2、β-catenin、PARP表达有关。
Abstract:
Objective To explore the effect and mechanism of deguelin on the proliferation and apoptosis of acute myeloid leukemia (AML) KG-1a cells.Methods The logarithmic growth phase KG-1a cells were randomly divided into 0.00 nmol·L-1 deguelin group,10.00 nmol·L-1 deguelin group,20.00 nmol·L-1 deguelin group,40.00 nmol·L-1 deguelin group and 80.00 nmol·L-1 deguelin group,and they were cultured with 0.00,10.00,20.00,40.00 and 80.00 nmol·L-1 deguelin respectively.After intervention with deguelin for 48 h,the morphological changes of cells in the five groups were observed with the inverted microscope.The proliferation ability of cells in the five groups were detected by the dimethylthiazole (MTT) experiment.In addition,KG-1a cells in the logarithmic growth phase were randomly divided into blank group,deguelin group,lithium chloride (LiCl) group,deguelin + LiCl group,the cells in the blank group were cultured in normal medium,the cells in the deguelin group were intervened with medium containing 80.00 nmol·L-1 deguelin for 72 h,the cells in the LiCl group were intervened with medium containing 30 mmol·L-1 LiCl for 72 h,the cells in the deguelin + LiCl group were intervened with medium containing 80.00 nmol·L-1 deguelin and 30 mmol·L-1 LiCl for 72 h.The apoptosis and cell cycle distribution of cells in the four groups were detected by flow cytometry.The relative expression levels of S-phase kinase-related protein 2 (SKP2),β-catenin and polyadenylic acid diphosphate ribose transferase (PARP) mRNA of cells in the four groups were detected by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) method.The relative expression levels of SKP2,β-catenin and PARP protein of cells in the four groups were detected by Western blot.Results After deguelin intervened for 48 h,compared with the 0.00 nmol·L-1 deguelin group,the cells in the 10.00 nmol·L-1 deguelin group,20.00 nmol·L-1 deguelin group,40.00 nmol·L-1 deguelin group and 80.00 nmol·L-1 deguelin group had larger intercellular spaces,significantly reduced number,irregular morphology,pyknosis of the nucleus and increased fragments.With the increase in the concentration of deguelin intervention,the number of cells gradually decreased,the morphological change was gradually obvious.After deguelin intervened for 24 h,48 h and 72 h,the cell proliferation capacity of cells in the 10.00 nmol·L-1 deguelin group,20.00 nmol·L-1 deguelin group,40.00 nmol·L-1 deguelin group and 80.00 nmol·L-1 deguelin group was significantly lower than that in the 0.00 nmol·L-1 deguelin group (P<0.05);the cell proliferation capacity of cells in the 20.00 nmol·L-1 deguelin group,40.00 nmol·L-1 deguelin group and 80.00 nmol·L-1 deguelin group was significantly lower than that in the 10.00 nmol·L-1 deguelin group (P<0.05);the cell proliferation capacity of cells in the 40.00 nmol·L-1 deguelin group and 80.00 nmol·L-1 deguelin group was significantly lower than that in the 20.00 nmol·L-1 deguelin group (P<0.05);the cell proliferation capacity of cells in the 80.00 nmol·L-1 deguelin group was significantly lower than that in the 40.00 nmol·L-1 deguelin group (P<0.05).After deguelin intervened for 24 h,48 h and 72 h,the proliferation ability of the cells in the five groups decreased with time,and the difference between each time point in the five groups was statistically significant (P<0.05).The apoptotic rate of cells in the blank group,deguelin group,LiCl group and deguelin+LiCl group was (5.37±0.61)%,(34.89±4.57)%,(2.54±0.36)%,(21.10±4.69)%,respectively;the apoptotic rate of cells in the blank group,LiCl group and deguelin+LiCl group was significantly lower than that in the deguelin group (P<0.05);the apoptotic rate of cells in the blank group and LiCl group was significantly lower than that in the deguelin+LiCl group(P<0.05);the apoptotic rate of cells in the LiCl group was significantly lower than that in the blank group (P<0.05).The proportion of cells in G0/G1 in the blank group,the deguelin + LiCl group and the deguelin group were significantly lower than that in the LiCl group,and the proportion of cells in G2/M was significantly higher than that in the LiCl group (P<0.05);the proportion of cells in G0/G1 in the deguelin + LiCl group and the deguelin group was significantly lower than that in the blank group,and proportion of cells in G2/M was significantly higher than that in the blank group (P<0.05);the proportion of cells in G0/G1 in the deguelin group was significantly lower than that in the deguelin + LiCl group,and proportion of cells in G2/M was significantly higher than that in the deguelin + LiCl group (P<0.05);there was no significant difference in the percentage of S-phase cells among the four groups (F=0.696,P>0.05).The relative expression levels of SKP2,β-catenin,PARP mRNA and protein in the blank group,deguelin + LiCl group and deguelin group were significantly lower than those in the LiCl group (P<0.05);the relative expression levels of SKP2,β-catenin,PARP mRNA and protein in the deguelin + LiCl group and deguelin group were significantly lower than those in the blank group (P<0.05);the relative expression levels of SKP2,β-catenin,PARP mRNA and protein in the deguelin group were significantly lower than those in the deguelin + LiCl group (P<0.05).Conclusion Deguelin has proliferation inhibition,cell cycle arrest and apoptosis promotion effects on AML KG-1a cells,and its mechanism of action may be related to the inhibition of SKP2,β-catenin and PARP expression.

参考文献/References:

[1] 王德征,江国虹,张爽,等.1999—2015年天津市白血病死亡率变化趋势分析[J].中华预防医学杂志,2019,53(3):319-322.
[2] BLACKBURN L M,BENDER S,BROWN S.Acute leukemia:diagnosis and treatment[J].Semin Oncol Nurs,2019,35(6):150950.
[3] MUKHERJEE S,SEKERES M A.Novel therapies in acute myeloid leukemia[J].Semin Oncol Nurs,2019,35(6):150955.
[4] LIAO W,LIU X,YANG Q,et al.Deguelin inhibits HCV replication through suppressing cellular autophagy via down regulation of Beclin1 expression in human hepatoma cells[J].Antiviral Res,2020,174:104704.
[5] NGUYEN C T,ANN J,SAHU R,et al.Discovery of novel anti-breast cancer agents derived from deguelin as inhibitors of heat shock protein 90 (HSP90)[J].Bioorg Med Chem Lett,2020,30(17):127374.
[6] GAO F,YU X,LI M,et al.Deguelin suppresses non-small cell lung cancer by inhibiting EGFR signaling and promoting GSK3β/FBW7-mediated Mcl-1 destabilization[J].Cell Death Dis,2020,11(2):143.
[7] 赵明明,黄艳群,李莲,等.Wnt/β-catenin信号通路激活剂对过氧化氢诱导的星形胶质细胞凋亡的影响[J].临床和实验医学杂志,2018,17(9):25-29.
[8] CHOPRA M,BOHLANDER S K.The cell of origin and the leukemia stem cell in acute myeloid leukemia[J].Genes Chromosomes Cancer,2019,58(12):850-858.
[9] VOSBERG S,GREIF P A.Clonal evolution of acute myeloid leukemia from diagnosis to relapse[J].Genes Chromosomes Cancer,2019,58(12):839-849.
[10] NARAYANAN D,WEINBERG O K.How I investigate acute myeloid leukemia[J].Int J Lab Hematol,2020,42(1):3-15.
[11] LI H,TIAN X,WANG P,et al.MicroRNA-582-3p negatively regulates cell proliferation and cell cycle progression in acute myeloid leukemia by targeting cyclin B2[J].Cell Mol Biol Lett,2019,4;24:66.
[12] CHEN L,JIANG K,CHEN H,et al.Deguelin induces apoptosis in colorectal cancer cells by activating the p38 MAPK pathway[J].Cancer Manag Res,2018,11:95-105.
[13] LI W,YU X,MA X,et al.Deguelin attenuates non-small cell lung cancer cell metastasis through inhibiting the CtsZ/FAK signaling pathway[J].Cell Signal,2018,50:131-141.
[14] ZHANG X,ZHAO Z,YI S,et al.Deguelin induced differentiation of mutated NPM1 acute myeloid leukemia in vivo and in vitro[J].Anticancer Drugs,2017,28(7):723-738.
[15] KANG Y A,PIETRAS E M,PASSEGU E.Deregulated Notch and Wnt signaling activates early-stage myeloid regeneration pathways in leukemia[J].J Exp Med,2020,217(3):jem.20190787.
[16] YUAN Y,WANG Q,MA S L,et al.lncRNA PCAT-1 interacting with FZD6 contributes to the malignancy of acute myeloid leukemia cells through activating Wnt/β-catenin signaling pathway[J].Am J Transl Res,2019,11(11):7104-7114.

相似文献/References:

[1]张婧婧,张楠,王侠,等.急性髓系白血病四色流式细胞术免疫学分型特点分析[J].新乡医学院学报,2011,28(03):323.
[2]崔珊珊,李雅慧,赵晨瑾,等.二甲双胍对人急性髓系白血病KG1a细胞增殖和凋亡的影响及其机制[J].新乡医学院学报,2021,38(10):901.[doi:10.7683/xxyxyxb.2021.10.001]
 CUI Shanshan,LI Yahui,ZHAO Chenjin,et al.Effect and mechanism of metformin on proliferation and apoptosis of human acute myeloid leukemia KG1a cells[J].Journal of Xinxiang Medical University,2021,38(12):901.[doi:10.7683/xxyxyxb.2021.10.001]
[3]张彦平,刘蒙蒙,展新荣.地西他滨联合小剂量高三尖杉酯碱和阿糖胞苷治疗老年急性髓系白血病疗效观察[J].新乡医学院学报,2020,37(11):1068.[doi:10.7683/xxyxyxb.2020.11.014]
 ZHANG Yanping,LIU Mengmeng,ZHAN Xinrong.Effect of decitabine combined with low-dose homoharringtonine and cytarabine in the treatment of elderly patients with acute myeloid leukemia[J].Journal of Xinxiang Medical University,2020,37(12):1068.[doi:10.7683/xxyxyxb.2020.11.014]
[4]申清云,高 大.老年急性髓系白血病的治疗方法研究进展[J].新乡医学院学报,2019,36(1):096.[doi:10.7683/xxyxyxb.2019.01.022]
[5]张彦平,刘蒙蒙,展新荣.地西他滨联合三氧化二砷治疗老年急性髓系白血病的疗效及安全性[J].新乡医学院学报,2019,36(12):1163.[doi:10.7683/xxyxyxb.2019.12.015]
 ZHANG Yan-ping,LIU Meng-meng,ZHAN Xin-rong.Evaluation of the efficacy and safety of decitabine combined with arsenic trioxide in the treatment of senile acute myeloid leukemia[J].Journal of Xinxiang Medical University,2019,36(12):1163.[doi:10.7683/xxyxyxb.2019.12.015]
[6]王继芳,魏秀丽.氟达拉滨联合阿糖胞苷治疗急性髓系白血病疗效观察[J].新乡医学院学报,2018,35(3):204.[doi:10.7683/xxyxyxb.2018.03.011]
 WANG Ji-fang,WEI Xiu-li.Effect of fludarabine combined with cytarabine on acute myeloid leukemia[J].Journal of Xinxiang Medical University,2018,35(12):204.[doi:10.7683/xxyxyxb.2018.03.011]
[7]尹俊杰,梁 波,王继芳,等.阿柔比星为基础的联合化学治疗方案治疗成人初治急性髓系白血病疗效观察[J].新乡医学院学报,2017,34(1):072.[doi:10.7683/xxyxyxb.2017.01.021]
 YIN Jun-jie,LIANG Bo,WANG Ji-fang,et al.Efficacy of aclarubicin-based chemotherapy induction regimens for adult de-novo acute myeloid leukemia[J].Journal of Xinxiang Medical University,2017,34(12):072.[doi:10.7683/xxyxyxb.2017.01.021]
[8]孟君霞,武永强,唐 广,等.CD7抗原表达与急性髓系白血病临床特征、细胞遗传学特点及预后的关系[J].新乡医学院学报,2016,33(6):508.[doi:10.7683/xxyxyxb.2016.06.017]
 MENG Jun-xia,WU Yong-qiang,TANG Guang,et al.Relationship between CD7 antigen expression and clinical features,cytogenetic features,prognosis of acute myeloid leukemia[J].Journal of Xinxiang Medical University,2016,33(12):508.[doi:10.7683/xxyxyxb.2016.06.017]
[9]张晓冬,林晓媛.急性髓系白血病患者外周血调节性T细胞、辅助性T细胞17及其相关细胞因子的表达及意义[J].新乡医学院学报,2023,40(4):339.[doi:10.7683/xxyxyxb.2023.04.008]
 ZHANG Xiaodong,LIN Xiaoyuan.Expression and significance of regulatory T cells,helper T cell 17 cells and their related cytokines in peripheral blood of patients with acute myeloid leukemia[J].Journal of Xinxiang Medical University,2023,40(12):339.[doi:10.7683/xxyxyxb.2023.04.008]

更新日期/Last Update: 2021-12-05