[1]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):901-905.[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(10):901-905.[doi:10.7683/xxyxyxb.2021.10.001]
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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
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
38
期数:
2021年10
页码:
901-905
栏目:
基础研究
出版日期:
2021-10-05

文章信息/Info

Title:
Effect and mechanism of metformin on proliferation and apoptosis of human acute myeloid leukemia KG1a cells
作者:
崔珊珊; 李雅慧; 赵晨瑾; 张　震; 姬曼曼; 王桐桐; 司晓慧; 牛新清
（新乡医学院医学检验学院，河南　新乡　453003）
Author(s):
CUI Shanshan; LI Yahui; ZHAO Chenjin; ZHANG Zhen; JI Manman; WANG Tongtong; SI Xiaohui; NIU Xinqing
(School of Laboratory Medicine,Xinxiang Medical University,Xinxiang 453003,Henan Province,China)
关键词:
二甲双胍; 急性髓系白血病; 细胞凋亡; 细胞增殖
Keywords:
metformin; acute myeloid leukemia; cell apoptosis; cell proliferation
分类号:
R733.7
DOI:
10.7683/xxyxyxb.2021.10.001
文献标志码:
A
摘要:
目的　探讨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。方法　取处于对数生长期KG1a细胞，用含体积分数10%胎牛血清的RPMI 1640培养基调整细胞悬液至终浓度为8×107 L-1，以每孔100 μL细胞悬液接种到96孔板中，随机分为对照组及2、4、6、8、10、12、16、24、32 mmol·L-1二甲双胍组，分别加入终浓度0、2、4、6、8、10、12、16、24、32 mmol·L-1二甲双胍，使用细胞计数试剂盒检测细胞存活率。另取处于对数生长期KG1a细胞，随机分为对照组、2 mmol·L-1二甲双胍组和6 mmol·L-1二甲双胍组，分别加入终浓度0、2 、6 mmol·L-1二甲双胍干预KG1a细胞，采用流式细胞术检测KG1a细胞凋亡率，免疫印迹法检测KG1a细胞中腺苷酸活化蛋白激酶α（AMPKα）、蛋白激酶B (AKT)、核糖体蛋白S6激酶（P70S6K）、4E-结合蛋白-1（4EBP-1）、Thr172位点磷酸化腺苷酸活化蛋白激酶［pAMPK(Thr172)］、Ser473位点磷酸化蛋白激酶B［pAKT（Ser473）］、Thr37/46位点磷酸化核糖体蛋白S6激酶［pP70S6K（Thr37/46）］、Thr389位点磷酸化4E-结合蛋白-1 ［p4EBP-1（Thr389）］相对表达量。结果　与对照组比较，2、4、6、8、10、12、16、24、32 mmol·L-1二甲双胍组KG1a细胞存活率显著减低（P＜0.05）；且各浓度二甲双胍组KG1a细胞的存活率呈浓度依赖性下降（P＜0.05）；二甲双胍对KG1a细胞的半数抑制浓度为6 mmol·L-1。2 mmol·L-1二甲双胍组和6 mmol·L-1二甲双胍组KG1a细胞凋亡率高于对照组（P＜0.05），6 mmol·L-1二甲双胍组KG1a细胞的凋亡率高于2 mmol·L-1二甲双胍组（P＜0.05）。2 mmol·L-1二甲双胍组和6 mmol·L-1二甲双胍组KG1a细胞中AKT、AMPKα、P70S6K、4EBP-1及P70S6K（Thr37/46）、p-4EBP-1（Thr389p）蛋白相对表达量显著低于对照组（P＜0.05），AMPKα（Thr172p）、p-AKT（Ser473）蛋白相对表达量显著高于对照组（P＜0.05）。6 mmol·L-1二甲双胍组KG1a细胞中AKT、AMPKα、P70S6K、4EBP-1及p-P70S6K（Thr37/46）、p-4EBP-1（Thr389p）蛋白相对表达量显著低于2 mmol·L-1二甲双胍组（P＜0.05），p-AMPKα（Thr172）、p-AKT（Ser473）蛋白相对表达量显著高于2 mmol·L-1二甲双胍组（P＜0.05）。结论　二甲双胍可抑制急性髓系白血病KG1a 细胞增殖并诱导其凋亡，其机制可能与二甲双胍调控AMPK信号通路有关。
Abstract:
Objective　To investigate the effect of metformin on the proliferation and apoptosis of human acute myeloid leukemia KG1a cells and its mechanism.Methods　KG1a cells in logarithmic growth stage were taken,and the cell suspension was adjusted to the final concentration of 8 × 107 L-1with RPMI1640 medium containing 10% fetal bovine serum,100 μL cell suspension was inoculated into 96 well plates,and KG1a cells was randomly divided into control group and 2,4,6,8,10,12,16,24,32 mmol·L-1 metformin group,the final concentration of 0,2,4,6,8,10,12,16,24 and 32 mmol·L-1 metformin was added,respectively;the cell survival rate was detected by cell counting kit.KG1a cells in logarithmic growth stage were randomly divided into the control group,2 mmol·L-1 metformin group and 6 mmol·L-1 metformin group,the final concentrations of 0,2 and 6 mmol·L-1 metformin were used to intervene KG1a cells,respectively.The apoptosis rate of KG1a cells was detected by flow cytometry,and the relative expressions of AMP-activated protein kinase α(AMPKα),protein kinase B(AKT),ribosomal protein S6 kinase(P70S6K）,eIF4E-binding proteins-1(4EBP-1),phosphorylated AMP-activated protein kinase at Thr172 site［pAMPK(Thr172)］,phosphorylated protein kinase B at Ser473 site［pAKT(Ser473)］,phosphorylated ribosomal protein S6 kinase at Thr37/46 site ［pP70S6K (Thr37/46)］,phosphorylated eIF4E-binding proteins-1 at Thr389 site ［p4EBP-1(Thr389)］ were detected by Western blot.Results　Compared with the control group,the survival rates of KG1a cells in 2,4,6,8,10,12,16,24,32 mmol·L-1 metformin group were decreased significantly (P＜0.05);the cell survival rate decreased significantly with the increase of metformin concentration (P＜0.05);the half inhibitory concentration of metformin on KG1a cells was 6 mmol·L-1.The apoptosis rate of KG1a cells in the 2 mmol·L-1 metformin group and 6 mmol·L-1 metformin group was significantly higher than that in the control group,the apoptosis rate of KG1a cells in the 6 mmol·L-1 metformin group was significantly higher than that in the 2 mmol·L-1 metformin group (P＜0.05).The relative expression levels of AKT,AMPKα,P70S6K,4EBP-1,P70S6K(Thr37/46),p-4EBP-1(Thr389) of KG1a cells in the 2 mmol·L-1 metformin group and 6 mmol·L-1 metformin group were significantly lower than those in the control group,the relative expression levels of p-AMPKα(Thr172) and p-AKT(Ser473) were significantly higher than those in the control group (P＜0.05).The relative expression levels of AKT,AMPKα,P70S6K,4EBP-1,p-P70S6K(Thr37/46),p-4EBP-1(Thr389) of KG1a cells in 6 mmol·L-1 metformin group were significantly lower than those in the 2 mmol·L-1 metformin group,and the relative expression levels of p-AMPKα(Thr172) and p-AKT(Ser473) protein were significantly higher than those in the 2 mmol·L-1 metformin group (P＜0.05).Conclusion　Metformin can inhibit the proliferation of acute myeloid leukemia KG1a cells and induce their apoptosis.The mechanism may be related to regulate the AMPK pathway.

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更新日期/Last Update: 2021-10-05