摘要
目的探究青藤碱(sinomenine,SIN)对人乳腺癌MCF-7细胞他莫昔芬(tamoxifen,TAM)耐药的逆转作用及相关机制。方法采用高剂量短时间刺激诱导法构建MCF-7/TAM耐药株,并通过MTT法验证细胞耐药性的改变,进一步采用MTT法检测SIN和TAM对MCF-7/TAM细胞的增殖抑制作用,并采用CompuSyn软件评估两者的联合效应。流式细胞术检测SIN和TAM对MCF-7/TAM细胞凋亡和周期的影响,Western blotting检测SIN对MCF-7/TAM细胞中PI3K/AKT/mTOR信号通路的影响。结果MCF-7/TAM细胞对TAM敏感性显著降低,已产生耐药性;SIN和TAM单用均能够抑制MCF-7/TAM细胞的增殖,TAM和SIN联合使用对MCF-7/TAM细胞的抑制作用显著增强,经CompuSyn软件分析显示两药具有协同作用;SIN和TAM联用能够更显著地诱导MCF-7/TAM细胞凋亡和阻滞细胞周期;MCF-7/TAM细胞中PI3K、Akt和mTOR的磷酸化水平显著高于MCF-7细胞,SIN干预后MCF-7/TAM细胞中PI3K、Akt和mTOR的磷酸化水平显著下降。结论SIN能够通过调控PI3K/AKT/mTOR信号通路,从而逆转MCF-7细胞对TAM耐药。
OBJECTIVE To investigate the mechanism of sinomenine(SIN)reversing drug resistance of human breast cancer MCF-7 cells to tamoxifen(TAM).METHODS MCF-7/TAM persister was constructed at high concentration for a short time by stimulation induction,and the changes of drug resistance were detected by MTT assay.The proliferation inhibition of SIN and TAM on MCF-7/TAM cells was further determined by MTT assay,and the combination index of the two drugs combined was analyzed by CompuSyn software.Flow cytometry was used to detect the effects of the two drugs on the apoptosis and cell cycle of MCF-7/TAM cells;the effect of SIN on the PI3K/AKT/mTOR signaling pathway in MCF-7/TAM cells was detected by Western blotting.RESULTS The sensitivity of MCF-7/TAM cell line to TAM was significantly reduced,and the drug resistance model was successfully constructed.Both SIN and TAM alone could inhibit the proliferation of MCF-7/TAM cells,and the inhibitory effect of TAM combined with SIN on MCF-7/TAM cells was significantly enhanced.CompuSyn software analysis showed that the two drugs were synergistic.Phosphorylation levels of PI3K,Akt and mTOR in MCF-7/TAM cells were significantly higher than those in MCF-7 cells.After SIN intervention,phosphorylation levels of PI3K,Akt and mTOR in MCF-7/TAM cells were significantly reduced.CONCLUSION Sinomenine can effectively reverse the resistance of MCF-7 cells to tamoxifen,possibly by regulating the PI3K/AKT/mTOR signaling pathway.
作者
俞萍萍
邵锦晖
YU Pingping;SHAO Jinhui(Hwa Mei Hospital,University of Chinese Academy of Sciences,Ningbo 315010,China;Hangzhou Fuyang District Hospital of Traditional Chinese Medicine,Hangzhou 311400,China)
出处
《中国现代应用药学》
CAS
CSCD
北大核心
2020年第24期2972-2978,共7页
Chinese Journal of Modern Applied Pharmacy
基金
浙江省自然科学基金项目(LY17H160061)。
关键词
青藤碱
乳腺癌
他莫昔芬
耐药
sinomenine
breast cancer
tamoxifen
drug resistance
