摘要
目的 观察哺乳动物雷帕霉素靶蛋白（mTOR）和磷酸肌醇3激酶（PI3K）特异性抑制剂RAD001和LY294002对人乳腺癌MCF-7、MDA-MB-231细胞株增殖与凋亡的影响,探讨PI3 K/蛋白激酶B （Akt）/mTOR信号通路上下游不同靶点联合阻滞对不同亚型人乳腺癌细胞株的抗肿瘤效应及其机制.方法 体外RAD001和LY294002单独或者联合使用,通过噻唑蓝（MTT）实验计算药物半数抑制剂量（IC50）.应用流式细胞仪分析细胞周期分布及凋亡.结果 LY294002和RAD001单药均使能人乳腺癌MCF-7和MDA-MB-231抑制细胞增殖、阻滞周期、诱导细胞凋亡.联合用药后各细胞株的抗肿瘤效应明显增强.结论 针对P13 K/Akt/mTOR信号通路的靶向干预可显著抑制人乳腺癌细胞的增殖,阻滞细胞周期并诱导其凋亡,且通路中不同靶点联合干预的抗肿瘤效应更加显著,尤其是雌激素受体（ER）、孕激素受体（PR）和人类表皮生长因子受体2（Her-2）均阴性的MDA-MB-231细胞株.
Objective In this study,by using LY294002 and RAD001,the specific inhibitors of phosphatidylinositol 3 kinase （PI3K） and mammalian target of rapamycin （mTOR）,to observe whether the anti-tumor effects on the proliferation,cycle distribution and apoptosis of combined inhibitors were a synergistic effect,and were differences between different molecular characteristics of human breast cancer cells alone and in combination in vitro.Methods Routinely cultured MCF-7 and MDA-MB-231 cells in vitro.Logarithmic phase cells among each cells were selected and divided into the blank control group,LY294002 group,RAD001 group and the combination group.Methyl thiazol tetrazolium （MTT） assay and flow cytometry were used to detect the cell proliferation,cell cycle distribution and cell apoptosis of different groups.Results LY294002 and RAD001 monotherapy were so capable breast cancer MCF-7 and MDA-MB-231 inhibited cell proliferation,cycle arrest,induction of apoptosis.This combination withered after each cell line significantly enhanced anti-tumor effect.Conclusion Targeted therapy on P13K/protein kinase B （Akt）/mTOR pathway could significantly inhibit the proliferation of human breast cancer cell lines by inducing apoptosis and arresting cell cycle distribution.By using a combination of different inhibitors which were targeted on different related genes of this pathway such as PI3K and mTOR,the anti-tumor effects were more significant increasd compared with monotherapy,especially in MDA-MB-231 cell line which were negative with ER,PR and human epidermal growth factor receptor （Her-2）.It was important to provide new research ideas for individualized treatment and translational medicine of breast cancer.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2015年第9期2143-2145,共3页
Chinese Journal of Experimental Surgery
