松果菊苷通过调节AKR1B10/ERK信号传导抑制乳腺癌MCF-7细胞的恶性进程  

Echinacoside hampers malignant progression of breast cancer MCF-7 cells by modulating AKR1B10/ERK signal transduction

作　　者：王倩婷 江砚 徐良辉 陈佳丽 WANG Qian-ting;JIANG Yan;XU Liang-hui;CHEN Jia-li(Pharmacy of Traditional Medicine,Affiliated Hangzhou First People's Hospital,School of Medicine,Zhejiang University,Hangzhou 310006,China)

机构地区：[1]浙江大学医学院附属杭州市第一人民医院中药房,浙江杭州310006

出　　处：《中国中药杂志》2023年第3期744-751,共8页China Journal of Chinese Materia Medica

基　　金：浙江省中医药科学研究基金项目(2022ZB274)。

摘　　要：分析松果菊苷(echinacoside)调控醛酮还原酶1B10(AKR1B10)/细胞外调节蛋白激酶(ERK)通路对乳腺癌(breast cancer,BC)MCF-7细胞增殖、转移以及阿霉素(adriamycin,ADR)耐药的作用。首先确定松果菊苷的化学结构,不同质量浓度(0、10、20、40μg·mL-1)松果菊苷对MCF-7细胞进行48 h干预,应用免疫印迹(Western blot)法分析AKR1B10/ERK通路相关蛋白的表达;细胞计数试剂盒(CCK-8)分析细胞活性。收集MCF-7细胞并分为对照组(control)、松果菊苷组(echinacoside)、松果菊苷+过表达对照组(echinacoside+Ov-NC)以及松果菊苷+AKR1B10过表达组(echinacoside+Ov-AKR1B10),Wes-tern blot分析AKR1B10/ERK通路相关蛋白的表达;CCK-8和EdU法分析细胞活性;划痕、Transwell实验、Western blot评估细胞转移。最后利用ADR对MCF-7细胞进行48 h干预,构建ADR耐药细胞,CCK-8检测细胞活性;原位末端标记法(TUNEL)和Western blot评估细胞凋亡。蛋白质资料库(PDB)数据库和分子对接预测了松果菊苷和AKR1B10两者的结合关系。与对照组相较,不同浓度松果菊苷可剂量依赖性削减MCF-7细胞中AKR1B10/ERK通路相关蛋白水平,细胞活性也表现出显著的下降趋势。与control比较,暴露于40μg·mL-1松果菊苷下的MCF-7细胞中AKR1B10/ERK通路、细胞增殖、转移和ADR耐药性被抑制;与echinacoside+Ov-NC相较,在松果菊苷处理后的MCF-7细胞中过表达AKR1B10会导致MCF-7细胞以上生物学行为部分恢复。松果菊苷与AKR1B10具有靶向关系。松果菊苷可阻碍乳腺癌细胞增殖、转移和ADR耐药,其机制可能与阻断AKR1B10/ERK通路有关。This study analyzes the impact of echinacoside(ECH) in the proliferation, metastasis and adriamycin(ADR) resistance of breast cancer(BC) MCF-7 cells via the modulation of aldo-keto reductase family 1 member 10(AKR1B10)/extracellular signal-regulated kinase(ERK) pathway. The chemical structure of ECH was firstly confirmed. MCF-7 cells were treated with different concentration(0, 10, 20, 40 μg·mL-1) of ECH for 48 h. Western blot was used to analyze expression of AKR1B10/ERK pathway-associated proteins and cell counting kit-8(CCK-8) assay to determine cell viability. MCF-7 cells were collected and classified into control group, ECH group, ECH + Ov-NC group, and ECH + Ov-AKR1B10 group. Then Western blot was employed to analyze the expression of AKR1B10/ERK pathway-associated proteins. CCK-8 and 5-ethynyl-2′-deoxyuridine(EdU) assay were used to examine cell proliferation. Cell migration was appraised with scratch assay, Transwell assay, and Western blot. Eventually, MCF-7 cells were treated with ADR for 48 h to induce ADR resistance. Cell viability was tested by CCK-8 assay and cell apoptosis was estimated based on terminal-deoxynucleoitidyl transferase mediated nick end labeling(TUNEL) assay and Western blot. Based on Protein Data Bank(PDB) and molecular docking, the binding affinity of ECH to AKR1B10 was assessed. Various doses of ECH decreased the expression of AKR1B10/ERK pathway-associated proteins in a dose-dependent manner and declined cell viability compared with the control group. Compared with the control group, 40 μg·mL-1ECH blocked the AKR1B10/ERK pathway in MCF-7 cells and inhibited the proliferation, metastasis and ADR resistance of the cells. Compared with the ECH + Ov-NC group, ECH + Ov-AKR1B10 group showed the recovery of some biological behaviors of MCF-7 cells. ECH also targeted AKR1B10. ECH can inhibit the proliferation, metastasis, and ADR resistance of BC cells by blocking AKR1B10/ERK pathway.

关 键 词：松果菊苷 AKR1B10/ERK信号 乳腺癌 阿霉素耐药 

分 类 号：R285[医药卫生—中药学]