AKBA联合阿霉素抑制三阴性乳腺癌细胞MDA-MB-231的增殖、迁移和裸鼠移植瘤生长  

AKBA combined with doxorubicin inhibits proliferation and metastasis of triple-negative breast cancer MDA-MB-231 cells and xenograft growth in nude mice

作　　者：曾佑琴[1] 陈思雨 刘燕 刘奕彤 张玲 夏姣 吴心语 魏常友 冷平[1] ZENG Youqin;CHEN Siyu;LIU Yan;LIU Yitong;ZHANG Ling;XIA Jiao;WU Xinyu;WEI Changyou;LENG Ping(College of Medical Technology,Chengdu University of Traditional Chinese Medicine,Chengdu 611130,China;School of Public Health,Chengdu Medical College,Chengdu 610500,China)

机构地区：[1]成都中医药大学医学技术学院,四川成都611130 [2]成都医学院公共卫生学院,四川成都610500

出　　处：《南方医科大学学报》2024年第12期2449-2460,共12页Journal of Southern Medical University

基　　金：四川省中医药管理局科研项目(2024MS563)。

摘　　要：目的探究AKBA联合阿霉素对三阴性乳腺癌细胞MDA-MB-231在增殖、迁移、侵袭和凋亡上的协同抑制作用,并通过网络药理学分析AKBA作用乳腺癌的下游信号通路。方法MDA-MB-231细胞体外培养,CCK-8法分别检测AKBA和阿霉素(ADR)作用MDA-MB-231细胞48 h的半抑制浓度(IC50),SynergyFinder在线网站(https://synergyfinder.fimm.fi)分析AKBA联合阿霉素的协同指数和最佳协同浓度。设置实验分组,对照组:MDA-MB-231正常培养;AKBA组:MDA-MB-231+AKBA(22.5μmol/L)处理48 h;ADR组:MDA-MB-231+(0.84μmol/L)处理48 h;AKBA+ADR组:MDA-MB-231+AKBA(22.5μmol/L)+ADR(0.84μmol/L)处理48 h,使用克隆形成实验、Transwell迁移和侵袭、划痕实验检测AKBA联合阿霉素抑制细胞迁移和侵袭的能力,采用Western blotting和qPCR实验检测凋亡相关基因的表达情况,将4~5周龄Balb/c nude小鼠随机分为4组(6只/组),建立裸鼠异种移植瘤模型,对照组:腹腔注射200μL 0.9%生理盐水;AKBA组:腹腔注射200μL50 mg/kg的AKBA;ADR组:腹腔注射200μL 2.5 mg/kg的ADR;AKBA+ADR组:腹腔注射200μL50 mg/kg的AKBA和2.5 mg/kg的ADR等量混合溶液,检测各组抑瘤率,HE染色检测组织病理情况,并通过网络药理学分析预测AKBA作用于乳腺癌的下游靶点和信号通路。结果AKBA对MDA-MB-231细胞作用48 h的IC50为45.15±0.97μmol/L,ADR的IC50为0.42±0.99μmol/L;SynergyFinder证实AKBA与ADR联合具有协同作用(ZIP>10);相较于对照组、AKBA组和阿霉素组,AKBA+ADR组明显抑制TNBC细胞增殖活性(P<0.0001),集落形成数量减少更多(P<0.05);相较对照组、AKBA和ADR组,AKBA+ADR组明显减弱细胞划痕愈合率(P<0.01);较对照组、AKBA组和ADR组,AKBA+ADR组所致细胞迁移至下室的数量减少更多(P<0.01);与对照组和AKBA或ADR单独使用相比,AKBA+ADR组抑制MDA-MB-231细胞侵袭基质胶能力增强(P<0.05);较AKBA或ADR单独作用,AKBA+ADR组在mRNA和蛋白水平上调Bax、Caspase-3剪切体和下调Bcl-2的表达更多(P<0.05);AKBA组、ADR�Objective To investigate the synergistic inhibitory effects of AKBA and doxorubicin on malignant phenotype of triple-negative breast cancer(TNBC)MDA-MB-231 cells.Methods CCK-8 assay was used to determine the 48-h IC50 of AKBA and doxorubicin in MDA-MB-231 cells,and SynergyFinder was employed to calculate the synergistic index and the optimal concentrations of the two agents.MDA-MB-231 cells treated with AKBA(22.5μmol/L),doxorubicin(0.84μmol/L)or their combination were examined for changes in cell proliferation,migration,invasion and apoptosis using Transwell migration,scratch assay,clone generation,RT-qPCR and Western blotting.Network pharmacology analysis was conducted to identify the downstream targets of AKBA in TNBC.In nude mouse models bearing subcutaneous MDA-MB-231 cell xenografts,the effects of normal saline,AKBA(50 mg/kg),doxorubicin(2.5 mg/kg),and AKBA combined with doxorubicin on xenograft growth and histopathology were observed.Results The IC50 of AKBA and doxorubicin in MDA-MB-231 cells at 48 h was 45.15±0.97μmol/L and 0.42±0.99μmol/L,respectively.SynergyFinder confirmed the synergistic effect of AKBA and ADR with a ZIP>10.The combined treatment with AKBA and doxorubicin significantly inhibited the proliferation,migration and invasion,promoted apoptosis of MDA-MB-231 cells,and effectively suppressed xenograft growth in nude mice.Network pharmacology analysis predicted that AKBA affects the progression of TNBC through its downstream target AKBA.Conclusion AKBA combined with doxorubicin inhibits proliferation,migration and invasion,promotes apoptosis of MDA-MB-231 cells and suppresses MDA-MB-231 cell xenograft growth in nude mice.The combined use of AKBA can attenuate the toxic effects of doxorubicin in nude mice.

关 键 词：AKBA 乙酰基-11-酮基-β-乳香酸 三阴性乳腺癌 阿霉素 

分 类 号：R737.9[医药卫生—肿瘤]