姜黄素联合西达本胺通过下调AKT磷酸化和上调p53表达调控SKM-1细胞增殖和凋亡  

Curcumin Combined with Chidamide Regulates Proliferation and Apoptosis of SKM-1 Cells by Suppressing AKT Phosphorylation and Upregulating p53 Expression

作　　者：邓琳丽 王利[1] DENG Linli;WANG Li(Department of Hematology,the First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学附属第一医院血液内科,重庆400016

出　　处：《中国细胞生物学学报》2021年第8期1614-1621,共8页Chinese Journal of Cell Biology

基　　金：重庆市教委基金(批准号:KJ1702017);重庆市科卫联合医学科研项目重点项目(批准号:2018ZDXM001);重庆市科委基础研究与前沿探索项目(批准号:cstc2018jcyjAX0688);重庆市渝中区基础研究与前沿探索项目(批准号:20190121)资助的课题。

摘　　要：该文探讨了姜黄素联合西达本胺对SKM-1细胞增殖和凋亡的影响及其作用机制。体外培养SKM-1细胞,取对数生长期细胞用于后续实验。对照组予以常规培养,实验组分别用不同浓度(1、5、10、20、40μmol/L)姜黄素、不同浓度(0.5、1、2、4、8μmol/L)西达本胺和不同浓度(5、10μmol/L)姜黄素联合不同浓度(0.5、1、2、4、8μmol/L)西达本胺处理细胞,采用CCK-8法检测各组细胞增殖活性,CompuSyn软件计算联合指数(combination index,CI),流式细胞术检测各组细胞周期分布和凋亡情况,Western blot检测各组细胞CDK2、p16、Caspase-3、AKT、p-AKT、p53和γH2A.X的蛋白表达水平。结果显示,在检测浓度范围内,姜黄素和西达本胺以时间浓度依赖性的方式抑制SKM-1细胞的生长。联合使用时,5μmol/L姜黄素与2μmol/L西达本胺具有协同抑制细胞增殖的作用。流式细胞术结果显示,5μmol/L姜黄素联合2μmol/L西达本胺组的细胞周期明显阻滞于G0/G1期,细胞凋亡率显著高于对照组和单独用药组。Western blot结果显示,与对照组相比,联合用药组的CDK2蛋白表达水平和p-AKT/AKT比例显著下降,而p16、Caspase-3、p53和γH2A.X的蛋白表达水平显著增高。综上,姜黄素联合西达本胺可显著抑制SKM-1细胞增殖,阻滞细胞周期,并促进细胞凋亡,其机制可能与抑制AKT磷酸化和上调p53表达有关。This study aimed to investigate the effect of curcumin combined with chidamide on SKM-1 cell proliferation and apoptosis and explore the underlying mechanism.SKM-1 cells were cultured in vitro,and the cells in the logarithmic growth phase were used for subsequent experiments.Cells in the control group were cultured routinely.Cells in the experimental groups were treated with different concentrations of curcumin(1,5,10,20,40μmol/L),different concentrations of chidamide(0.5,1,2,4,8μmol/L)and different concentrations of curcumin(5,10μmol/L)combined with different concentrations of chidamide(0.5,1,2,4,8μmol/L),respectively.CCK-8 assay was used to detect the proliferation activity.CI(combination index)was calculated by Compusyn software.Cell cycle distribution and apoptosis were detected by flow cytometry.The protein expression levels of CDK2,p16,Caspase-3,AKT,p-AKT,p53 andγH2A.X in each group were detected by Western blot.The results showed that curcumin and chidamide inhibited the growth of SKM-1 cells in a time-and concentration-dependent manner.Curcumin(5μmol/L)and chidamide(2μmol/L)had a synergistic inhibitory effect on cell proliferation.The results of flow cytometry showed that 5μmol/L curcumin combined with 2μmol/L chidamide arrested the cell cycle at G0/G1 phase,and the apoptotic rate in the combination treatment group was significantly higher than that in the control and the single drug groups.The results of Western blot showed that compared with the control group,in the combination treatment group,the protein expression level of CDK2 and p-AKT/AKT ratio were significantly decreased while the protein expression levels of p16,caspase-3,p53 andγH2A.X were significantly increased.Collectively,curcumin combined with chidamide could significantly inhibit the proliferation of SKM-1 cells,block cell cycle and promote cell apoptosis,whose mechanism might be related to the inhibition of AKT phosphorylation and upregulation of p53 expression.

关 键 词：姜黄素 西达本胺 SKM-1细胞 AKT P53 细胞增殖 

分 类 号：R285[医药卫生—中药学]