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

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/mTOR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?mL-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?mL-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/mTOR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?mL-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-mTOR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-mTOR和p-Akt的活性，提高了PTEN的表达有关。

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

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/mTOR 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 ? mL-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 ? mL-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/ mTOR 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 ? mL-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-mTOR 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-mTOR and p-Akt, and also up-regulating the expressions of PTEN.