【摘要】 目的通过联合抑制MEK和AKT信号通路关键靶点分子,探讨克服细胞耐药性的靶向治疗方法。方法体外培养和新鲜分离恶性黑色素瘤细胞株,分别在0.5%和5%血清浓度下单独或同时抑制靶点MEK(U0126,20μmol/L)和AKT(LY294002,20μmol/L),MTT法检测细胞增殖,PI染色流式细胞术检测细胞凋亡情况。结果当培养液血清浓度从0.5%提高到5%时,U0126和LY294002对肿瘤细胞增殖的抑制效应明显降低。LY294002以抑制细胞增殖为主,U0126主要诱导细胞凋亡,二者作用效果与细胞是否具备该通路关键靶点活化变异无关。联合用药时诱导细胞凋亡普遍增强,LY294002和U0126具有协同效应。结论联合抑制MEK/ERK和PI3K/AKT信号通路可较好地克服恶性黑色素瘤细胞的耐药性。更多还原

【Abstract】 Objective The treatment of metastatic melanoma by conventional chemotherapeutic agents remains unsatisfactory.The present study was undertaken to reveal the role of co-inhibition of survival signaling pathways in apoptosis of melanoma cells.Methods A panel of human melanoma cell lines and fresh melanoma isolates was assessed for their sensitivity to the MEK inhibitor U0126and/or AKT inhibitor LY294002.The proliferation and apoptosis of the cells were examined after treatment with the inhibitors.Results Constitutive activation of ERK1/2and AKT was closely related to concentrations of serum in the culture medium(extracellular signals).The sensitivity of melanoma cells to apoptosis induced by inhibition of MEK/ERK was not correlated with the active BRAF mutation(BRAFV600E).Inhibition of MEK/ERK predominantly induced apoptosis;whereas inhibition of PI3K/AKT primarily inhibited proliferation.Co-inhibition of MEK/ERK1/2 and PI3K/AKT synergistically induced apoptosis.Conclusion Co-targeting MEK/ERK and PI3K/AKT pathways may further improve treatment for melanoma.更多还原