南蛇藤素通过调控PTEN/Akt/Raf-1信号通路增强乳腺癌细胞对环磷酰胺的化疗敏感性  

Celastrol Enhances Chemosensitivity of Conditionally Reprogrammed Breast Cancer Cells to Cyclophos-phamide through Modulation of PTEN/Akt/Raf-1 Signaling Pathway

作　　者：李嘉琪 石艺 钟燕 唐欲博[1] 林颖[2] 陈孝[1] LI Jiaqi;SHI Yi;ZHONG Yan;TANG Yubo;LIN Ying;CHEN Xiao(Department of Pharmacy,The First Affiliated Hospital of Sun Yat-Sen University,Guangzhou,Guangdong 510080,China;Thyroid&Breast Surgery,The First Affiliated Hospital of Sun Yat-Sen University,Guangzhou,Guangdong 510080,China)

机构地区：[1]中山大学附属第一医院药学部,广东广州510080 [2]中山大学附属第一医院甲状腺乳腺外科,广东广州510080

出　　处：《今日药学》2021年第4期255-262,共8页Pharmacy Today

基　　金：国家重点研发计划“精准医学研究”专项(2017YFC0909900);广东省基础与应用基础研究基金项目(2020B1515120094)。

摘　　要：目的探究南蛇藤素是否能增强环磷酰胺的抗乳腺癌作用及其机制。方法收集临床患者浸润性乳腺癌手术切除组织,利用条件性重编程细胞技术培养患者来源的乳腺癌细胞,并用Western Blot和免疫细胞化学检测乳腺癌细胞生物标记物抗癌胚抗原、雌激素受体和上皮细胞黏附分子表达,鉴定乳腺癌细胞表型。采用MTS检测不同浓度南蛇藤素和环磷酰胺及两者联合处理后细胞的增殖活力,Calcein-AM/PI和Hoechst33258/PI双染法用于检测药物处理后细胞活力及凋亡状态,transwell及Matrigel血管形成实验用于评估南蛇藤素和环磷酰胺对细胞迁移能力及肿瘤血管生成能力的作用,Western Blot检测细胞蛋白表达水平。结果南蛇藤素和环磷酰胺均可抑制细胞增殖且呈浓度依赖性,与单一药物处理相比,联合南蛇藤素和环磷酰胺可显著抑制细胞增殖、迁移,促进细胞凋亡,且明显抑制管腔形成,上调乳腺癌细胞中PTEN磷酸化水平而抑制Akt和Raf-1磷酸化。结论南蛇藤素可增强乳腺癌对环磷酰胺的化疗敏感性,其机制可能与激活PTEN并下调Akt/Raf-1通路蛋白磷酸化有关。OBJECTIVE To investigate whether celastrol can enhance the anti-breast cancer effect of cyclophosphamide and the underlying mechanism.METHODS Clinical tissues of invasive breast cancer were collected and patient-derived breast cancer cells were cultivated using conditional reprogramming technology.To identify cell phenotype,the expression of ER,CEA and EpCAM of the cells were detected by western blot and immunocytochemistry.MTS was used to detect the proliferation ability after cells were treated with different concentrations of celastrol or cyclophosphamide,or the combination of them.Calcein-AM/PI and Hoechst33258/PI dual-staining were used to estimate the viability and apoptosis respectively.Transwell assay and the capillary tube formation assay were used to evaluate cell migration and tube formation ability respectively.Western blot was used to detect protein expression of the cells.RESULTS Both celastrol and cyclophosphamide exhibited inhibitory effects on the cell proliferation in a concentration-dependent manner.Compared with single drug treatment,the combination of celastrol and cyclophosphamide inhibited cell proliferation and migration,promoted cell apoptosis,and inhibited capillary structure formation of human umbilical vein endothelial cells(HUVECs)significantly.Moreover,the phosphorylation level of PTEN was upregulated,while the phosphorylation of Akt and Raf-1 were downregulated by the combination of the two drugs.CONCLUSION Celastrol enhances the antitumor effects of cyclophosphamide against breast cancer cells.And the mechanism may be related to the activation of PTEN and phosphorylation down-regulating of Akt/Raf-1 signaling pathway.

关 键 词：南蛇藤素 环磷酰胺 条件性重编程乳腺癌细胞 细胞增殖 PTEN Akt/Raf-1信号通路 

分 类 号：R966[医药卫生—药理学]