益气扶正复方联合依维莫司对三阴性乳腺癌细胞株MDA-MB-468的增殖抑制作用及其对Akt/mTOR信号通路的影响  

Proliferation Inhibition Effect of Yiqi Fuzheng Formula Combined with Everolimus on Proliferation of Triple Negative Breast Cancer Cell Line MDA-MB-468 Cells and Its Impact on Akt/mTOR Signaling Pathway

作　　者：李甜 周钱梅[2] 张卫红[1] Li Tian;Zhou Qianmei;Zhang Weihong(Breast Surgery Department, Shuguang Hospital Baoshan Branch Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201900, China;Complex System Research Center Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China)

机构地区：[1]上海中医药大学附属曙光医院宝山分院乳腺科,上海201900 [2]上海中医药大学中医复杂系统研究中心,上海201203

出　　处：《世界科学技术-中医药现代化》2018年第1期37-43,共7页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：上海市卫生和计划生育委员会科研课题面上项目:基于雌激素受体基因多态性的补肾中药防治芳香化酶抑制剂(AIs)引起的骨代谢异常的临床研究(201540036);负责人:张卫红;国家自然科学基金青年科学基金项目(8150150222):益气扶正法对三阴性乳腺癌Akt/m TOR信号通路的影响;负责人:张卫红;上海市宝山区中西医结合医院国家自然培育基金项目(GZRPYJJ-201702):从自噬角度探讨芒柄花素对三阴性乳腺癌MDA-MB-231紫杉醇耐药细胞株的作用机制;负责人:张卫红

摘　　要：目的探讨益气扶正复方(简称复方)联合依维莫司对三阴性乳腺癌细胞株MDA-MB-468细胞的增殖抑制作用及Akt/m TOR信号通路的影响。方法高效液相色谱法(HPLC)制备益气扶正复方药物,以MTT比色法分别检测10,20,40,80,160 mg?m L-1复方药物及1,10,100,1000 nmol?L-1依维莫司(Everolimus)12,24,48 h对MDA-MB-468细胞的增殖抑制作用;分别运用IC50浓度的复方40 mg?m L-1与Everolimus 100 nmol?L-1单独及联合干预MDA-MB-468乳腺癌细胞48 h,观察其对细胞增殖的抑制作用,流式细胞仪检测其对细胞周期和凋亡的影响;Western blot法检测其对细胞Akt/m TOR信号通路的影响。结果复方及Everolimus均呈剂量、时间依赖性地抑制了MDA-MB-468乳腺癌细胞增殖,选择IC50浓度的复方40 mg?m L-1与Everolimus 100 nmol?L-1单独及联合干预细胞,联合用药组显著抑制了MDA-MB-468乳腺癌细胞的增殖,同时联合用药组抑制了p-m TOR和p-Akt的活性,提高了PTEN的表达。结论复方与Everolimus均能抑制MDA-MB-468乳腺癌细胞增殖,两药联用效果更为显著,其作用机制可能与复方与依维莫斯联用抑制了p-m TOR和p-Akt的活性,提高了PTEN的表达有关。This study was aimed to explore effects of Yiqi Fuzheng formula（YFF）joint with Everolimus on cell cycle and apoptosis of triple negative breast cancer（TNBC）cell line MDA-MB-468 cells,and to observe its impact on Akt/m TOR signal pathway.High performance liquid chromatography（HPLC）was used for the preparation of YFF.Methyl thiazolyl tetrazolium（MTT）assay was used to detect the MDA-MB-468 cell proliferation inhibition of 10,20,40,80,160 mg · m L^-1 YFF and 1,10,100,1000 nmol · L^-1 of Everolimus for 12,24 and 48 h.MDA-MB-468 breast cancer cells were treated with IC50 YFF of 40 mg · m L^-1 and Everolimus of 100 nmol · L^-1 for 48 h.And then,the inhibition of cell proliferation was observed and the cell cycle and apoptosis were detected by flow cytometry.Western blot was used to detect the effect of Akt/m TOR signal pathway on cells.The results showed that YFF and Everolimus inhibited the proliferation of MDA-MB-468 breast cancer cells in a dose-and time-dependent manner.Intervened individually and jointly by IC50 YFF of 40 mg · m L^-1 and Everolimus of 100 nmol · L^-1,the combined treatment group significantly inhibited the proliferation of MDA-MB-468 breast cancer cells.At the meantime,it inhibited the activity of p-m TOR and p-Akt and increased the expression of PTEN.It was concluded that both YFF and Everolimus can inhibit proliferation of TNBC cell line MDA-MB-468,while the combination group had more significant effect.The antiproliferation mechanisms may be related to its effects of repressing the activity of p-m TOR and p-Akt,and also up-regulating the expressions of PTEN.

关 键 词：三阴性乳腺癌(TNBC) 益气扶正复方(复方) 细胞增殖抑制 Akt/mTOR信号通路 

分 类 号：R33[医药卫生—人体生理学]