高三尖杉酯碱对人三阴性乳腺癌细胞迁移和侵袭能力的影响  

Effects of homoharringtonine on the migration and invasion ability of human triple negative breast cancer cells

作　　者：周蕾[1] 李雅丽 冷红 ZHOU Lei;LI Yai;LENG Hong(Thyroid Breast Surgery,Meishan People's Hospital,Meishan 620010,Sichuan,China)

机构地区：[1]眉山市人民医院甲乳外科,眉山620010

出　　处：《医学研究与战创伤救治》2024年第6期567-574,共8页Journal of Medical Research & Combat Trauma Care

基　　金：四川省医学(青年创新)科研课题(Q20168)。

摘　　要：目的 探讨高三尖杉酯碱(HHT)对人三阴性乳腺癌(TNBC)细胞系MDA-MB-231迁移和侵袭的影响,并初步分析潜在机制。方法 采用四甲基偶氮唑盐法(MTT法)检测细胞活力。采用EdU结合实验以评估细胞增殖。分别采用细胞划痕实验和Transwell实验检测细胞迁移与侵袭。采用流式细胞术分析乳腺癌干细胞(BCSC)含量。采用球体形成试验评估MDAMB-231细胞“肿瘤球”形成能力。采用实时定量聚合酶链式反应(qRT-PCR)检测Hedgehog(HH)信号通路效应因子Gli1 mRNA和乳腺癌细胞干性指标Oct4、CD44、Sox2和Nanog mRNA的表达。采用蛋白免疫印迹分析上皮间质转化(EMT)标志物E-cadherin、N-cadherin、Vimentin蛋白以及Gil1蛋白表达。结果 HHT以剂量依赖性的方式降低MDA-MB-231细胞活力(P<0.05)。HHT可抑制BC细胞的增殖、迁移和侵袭。此外,HHT可下调BC细胞中CD44+/CD24-细胞比例、降低MDA-MB-231细胞形成“肿瘤球”的能力并抑制Oct4、CD44、Sox2和Nanog mRNA表达,提示HHT可抑制MDA-MB-231细胞干性。HHT可通过抑制“肿瘤球”中E-cadherin表达并增加N-cadherin和Vimentin蛋白来可抑制BCSC的EMT。此外,HHT可抑制TNBC中HH/Gli1信号通路活化。同时,HHT能够消除Gli1过表达对TNBC干性和EMT的影响。结论 HHT能够抑制乳腺癌细胞的迁移和侵袭,这可能是通过调控HH/Gli1信号通路以抑制TNBC的干性和EMT来实现。Objective To investigate the effects of homoharringtonine(HHT)on migration and invasion of human triple nega-tive breast cancer(TNBC)cell line MDA-MB-231,and to analyze the potential mechanisms.Methods MTT assay was used to de-tect cell viability.EdU incorporation assay was used to evaluate cell proliferation.Cell scratch assay and Transwell assay were used to detect cell migration and invasion respectively.Flow cytometry was used to analyze the content of breast cancer stem cells(BCSC).Sphere formation test was used to evaluate the ability of MDA-MB-231 cells to form tumor spheres.Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect Gli1 mRNA and the mRNA expression of Oct4,CD44,Sox2 and Nanog in breast cancer cells.Western blotting was used to analyze the expression of EMT markers E-cadherin,N-cadherin,Vimentin and Gil1 protein.Results Homoharringtonine decreased the viability of MDA-MB-231 cells in a dose-dependent manner(P<0.05).Homoharringtonine could in-hibit the proliferation,migration and invasion of BC cells.In addition,homoharringtonine could down-regulate the ratio of CD44+/CD24-cells in BC cells,decreased the ability of MDA-MB-231 cells to form tumor balls,and inhibited the mRNA expressions of Oct4,CD44,Sox2 and Nanog,suggesting that homoharringtonine could inhibit the dryness of MDA-MB-231 cells.Homoharringtonine could inhibit EMT of BCSC by inhibiting E-cadherin expression and increasing N-cad-herin and Vimentin proteins in tumor spheres.In addition,homoharringtonine could inhibit the activation of HH/Gli1 signaling pathway in TNBC.Meanwhile,homoharringtonine could eliminate the effects of Gli1 overexpression on TNBC stemness and EMT.Conclu‐sion Homoharringtonine can inhibit the migration and invasion of breast cancer cells,which may be achieved by regulating HH/Gli1 signaling pathway to inhibit TNBC stemness and EMT.

关 键 词：高三尖杉酯碱 三阴性乳腺癌 迁移和侵袭 HH/Gli1信号通路 乳腺癌干细胞 上皮间质转化 

分 类 号：R737.9[医药卫生—肿瘤]