二甲双胍和雷帕霉素联用对MYCN扩增神经母细胞瘤细胞的作用

The Effect of Metformin Combined with Rapamycin on the Proliferation of MYCN-amplified Neuroblastoma Cells

目的探讨二甲双胍(metformin,Met)和雷帕霉素(rapamycin,RAPA)联用对MYCN扩增神经母细胞瘤细胞的作用及其机制。方法采用MTT法检测二甲双胍和雷帕霉素对MYCN扩增神经母细胞瘤细胞增殖的抑制作用,并判断两药联用时的效应;采用试剂盒检测培养基上清中的乳酸和葡萄糖含量的变化,Western blot法检测磷酸化AKT(Phosphorylation AKT,p-AKT)和磷酸化S6K(Phosphorylation S6K,p-S6K)蛋白表达的变化。结果二甲双胍和雷帕霉素联用对MYCN扩增神经母细胞瘤细胞的增殖具有明显的抑制作用,其两药合用指数(CI)<1;与单用二甲双胍组相比,联用组的葡萄糖吸收和乳酸分泌量明显减少( P <0.05);与单独用药组相比,联用组的磷酸化AKT和p-S6K蛋白表达水平均明显降低( P <0.05)。结论二甲双胍与雷帕霉素联用对MYCN扩增神经母细胞瘤细胞的抑制作用具有协同效应,其机制可能与降低乳酸的堆积和抑制AKT/mTOR信号通路有关。

Objective To investigate the effect of metformin combined with rapamycin on the proliferation of MYCN-amplified neuroblastoma cells and its mechanism. Methods MTT assay was used to detect the inhibitory effect of metformin and rapamycin on the proliferation of MYCN-amplified neuroblastoma cells and to determine the effect of the combination of the two drugs.The changes of lactic acid and glucose content in the supernatant of culture medium were detected by kit.The changes of phosphorylated AKT (P-AKT) and phosphorylated S6K (Phosphorylation S6K,p-S6K) protein expression were detected by Western blot. Results Metformin combined with rapamycin inhibited the proliferation of MYCN-amplified neuroblastoma cells,and the combined use index (CI) was less than one.Compared with metformin alone,the glucose absorption and lactate secretion in the combined use group were significantly reduced ( P <0.05),and the phosphorylated AKT and S6K protein expression levels in the combined use group were significantly lower than those in the single use group ( P <0.05). Conclusion Metformin combined with rapamycin has synergistic effect on the proliferation of MYCN-amplified neuroblastoma cells.The mechanism may be related to the reduction of lactate accumulation and the inhibition of AKT/mTOR signaling pathway.