Effect of oxaliplatin combined with AG1478 on lung cancer by regulating reactive oxygen species and inhibiting PI3K/AKT pathway
Objective To investigate the effect of epidermal growth factor tyrosine kinase inhibitor AG1478 combined with oxaliplatin(OXA)on non-small cell lung cancer cells H1299.Methods H1299 cells were divided into control group(conventional culture,without drug),OXA-25,-50,-100 groups(25,50 and 100 μmol·L-1 OXA),AG-20 group(20 μmol·L-1AG1478)and OXA+AG group(25 μmol·L-1 OXA and 20 μmol·L-1AG1478).Detection of cell survival rate by MTT assay and calculation of half inhibitory concentration(IC50).Reactive oxygen species(ROS)probe H2DCFDA was used to detect ROS levels.Autophagy of H1299 cells was detected by MDC method.Western blot was used to detect the expression levels of phosphorylated-phosphoinositide 3-kinase(p-PI3 K),phosphorylated-protein kinase B(p-AKT),autophagy protein(Beclin-1)and microtubule associated protein light chain 3-Ⅱ(LC3-Ⅱ).Results The IC50 of OXA on H1299 cells was 91.09 μmol·L-1.The IC50 of AG1478 on H1299 cells was 31.83 μmol·L-1.The relative ROS level were 1.00±0.03,1.15±0.02,1.76±0.04,2.89±0.02,1.05±0.01 and 3.20±0.03,respectively;the relative MDC levels were 1.00±0.04,1.10±0.02,1.16±0.02,1.46±0.04,1.04±0.01 and 1.31±0.02,respectively;the relative expression levels of p-PI3K protein were 1.12±0.05,0.88±0.06,0.72±0.07,0.60±0.05,0.91±0.07 and 0.64±0.09,respectively;the relative expression levels of p-AKT protein were 1.09±0.04,0.87±0.08,0.77±0.07,0.63±0.05,0.76±0.05 and 0.46±0.03,respectively;the relative expression levels of Beclin-1 protein were 0.82±0.03,0.91±0.04,1.06±0.28,1.11±0.03,0.87±0.04 and 1.27±0.10,respectively;the relative expression levels of LC3-Ⅱ protein were 0.65±0.08,0.82±0.11,1.08±0.12,1.38±0.09,0.72±0.11 and 1.38±0.15,respectively.Compared the OXA+AG group and the OXA single group with the control group,compared the OXA+AG group with the OXA single group,there were statistically significant differences in the above indicators(P＜0.05,P＜0.01).Conclusion AG1478 combined with OXA can increase ROS levels in non-small cell lung cancer cells H1299 and inhibit the activation of PI3K signaling pathway,thus inducing autophagy.
国家科技期刊平台
NSTL
万方数据
