摘要
目的基于自噬-NF-κB通路探讨阿魏酸钠对白血病化疗耐药株增敏的机制。方法复制白血病化疗耐药细胞模型,Western blot法观察L1210、L1210/DDP细胞自噬-NF-κB通路的基础表达水平,将L1210/DDP随机分为control组(L1210/DDP)、model组(L1210/DDP+8μg/ml DDP)、SF组(model组+0.5mmol/L SF)、3-MA组(model组+10μmol/L 3-MA)、SN50组(model组+2μmol/L SN50)、SF+3-MA组(model组+0.5mmol/L SF+10μmol/L 3-MA)及SF+SN50组(model组+0.5mmol/L SF+2μmol/L SN50)7组。据实验分组进行药物干预48h,行指标检测。cck-8法检测实验各组细胞活力,流式细胞仪检测实验各组细胞凋亡率、细胞周期的变化;Western blot检测实验各组细胞自噬-NF-κB通路关键节点蛋白表达变化。结果与L1210细胞相比,L1210/DDP细胞自噬与NF-κB通路异常激活(P<0.05);与model组相比,各给药组细胞活力降低、G0/G1期细胞百分比减少(P<0.05)、S期细胞百分比增加(P<0.05)、凋亡率增加(P<0.05),且SF与抑制剂联用效果更佳,LC3BⅡ/Ⅰ表达减少(P<0.05)、p-p65/p65比值降低、bcl-2/bax表达减少(P<0.05);与SF组相比,SF与抑制剂联用组Beclin1、LC3BⅡ/Ⅰ蛋白减少(P<0.05),bcl-2/bax比值降低(P<0.05)。结论SF通过抑制耐药株自噬-NF-κB通路,增加耐药株对DDP的敏感性,具有抑制癌细胞增殖,促进癌细胞凋亡的功效。
Objective To investigate the effect of Sodium Ferulate(SF)on chemosensitivity of leukemia-resistant based on the autophagy-NF-κB pathway.Methods Establish the leukemia chemotherapy-resistant cell model,observe the basic expression level of autophagy-NF-κB pathway between L1210 and L1210/DDP cells by Western blot,and L1210/DDP cells were randomly divided into 7 group:control group(L1210/DDP),model group(L1210/DDP+8μg/ml DDP),SF group(model group+0.5 mmol/L SF),3-MA group(model group+10μmol/L 3-MA),SN50 group(model group+2μmol/L SN50),SF+3-MA group(model group+0.5 mmol/L SF+10μmol/L 3-MA)and SF+SN50 group(model group+0.5 mmol/L SF+2μmol/L SN50).According to the experimental group,the treated group was dealed with the corresponding drug for 48 h,and the indicators were tested.The cell viability of each group was measured by the cck-8 method.The apoptosis rate and cell cycle changes of each experimental group were detected by flow cytometry.The expression changes of key proteins in each node of autophagy-NF-κB pathway were detected by Western blot.Results Compared with L1210 cells,the autophagy and NF-κB pathway were abnormally activated in the L1210/DDP cells(P<0.05).Compared with the model group,the cell viability and the percentage of G0/G1 phase decreased in the treated groups(P<0.05).Moreover,the percentage of S phase and apoptosis rate increased in the treated groups(P<0.05),and SF combined with inhibitors had the best effect.LC3 BII/Ⅰand bcl-2/bax decreased(P<0.05),and p-p65/p65 ratio was reduced in the treated groups.Compared with the SF group,the proteins express of Beclin1 and LC3 BII/Ⅰand bcl-2/bax ratio were reduced(P<0.05)in the SF combined with inhibitors group.Conclusion SF increases the sensitivity of drug-resistant strains to DDP by inhibiting the autophagy-NF-κB pathway,and shows an effective activity in inhibiting cell proliferation and promoting cell apoptosis.
作者
何欢
郑倩倩
曾雪倩
侯梦雪
王旭
王晓玲
汪涛
HE Huan;ZHENG Qian-qian;ZENG Xue-qian;HOU Meng-xue;WANG Xu;WANG Xiao-ling;WANG Tao(Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China)
出处
《时珍国医国药》
CAS
CSCD
北大核心
2021年第6期1297-1300,共4页
Lishizhen Medicine and Materia Medica Research
基金
国家自然科学基金(81503401,81673732)
天津自然科学基金项目(18JCYBJC27100)
大学生科技创新基金项目(CXJJ2020YF16)
研究生科研创新(YJSKC-20191032,ZXYCXLX201910)。
