奥希替尼联合槲皮素对H1975细胞增殖的协同抑制作用

Synergistic effect of osimertinib combined with quercetin inhibiting H1975 cells proliferation

目的 探索奥希替尼联合槲皮素对非小细胞肺癌(NSCLC)H1975细胞的增殖抑制作用以及潜在的作用机制。方法 采用MTT法检测2.625、5.25、10.5、21、42、84 nmol/L奥希替尼和15.625、31.25、62.5、125、250、500μmol/L槲皮素对细胞增殖的抑制效应(设置单一药物组及药物联合组),根据检测的结果计算得出奥希替尼和槲皮素对细胞的半数抑制浓度(IC50)以及联合指数(CI)。单药组、两药联合组及空白对照组的细胞凋亡率采用流式细胞仪进行检测。Western blot检测各组细胞中Akt/p-Akt、Bcl-2及Bax蛋白的表达水平变化。结果 不同浓度的奥希替尼和槲皮素均可抑制H1975细胞的增殖(P<0.01),IC50值分别为(21.54±1.05) nmol/L和(109.7±1.08)μmol/L;各浓度奥希替尼与槲皮素联合时的CI值均<1,表现为协同抑制效应。两药联合作用后表现出更强的诱导细胞凋亡作用。奥希替尼联合槲皮素组相对两药单药组可观察到p-Akt及Bcl-2蛋白的表达明显下调(P<0.01),而两药联合组Bax蛋白的表达水平显著高于单药组(P<0.01)。结论 奥希替尼联合槲皮素表现出的协同效应使H1975细胞增殖受到明显抑制,其机制可能与药物作用使p-Akt蛋白的表达下调以及促细胞凋亡有关。

Objective To explore the effect of osimertinib combined with quercetin inhibiting non-small cell lung cancer(NSCLC)H1975 cells proliferation and potential mechanism.Methods MTT method was used to detect the effects of osimertinib(2.625,5.25,10.5,21,42 and 84 nmol/L)and quercetin(15.625,31.25,62.5,125,250 and 500μmol/L)as single drug and the combination of the two drugs on proliferation of H1975 cells.The half inhibitory concentration(IC50)and the combination index(CI)values of osimertinib and quercetin on H1975 cells were calculated.The apoptotic rate of each group of H1975 cells was detected by flow cytometry.The changes of Akt/p-Akt,bcl-2 and bax protein expression levels in cells of each group were tested by Western blot.Results Osimertinib and quercetin at different concentrations could inhibit the proliferation of H1975 cells(both P<0.01).The IC50 values were(21.54±1.05)nmol/L and(109.7±1.08)μmol/L,respectively.CI values were<1 when osimertinib combined with quercetin at all concentrations,showing a synergistic inhibitory effect.The combination of the two drugs showed a stronger effect of inducing apoptosis.The expressions of p-Akt and bcl-2 proteins in the osimertinib combined with quercetin group were significantly down-regulated compared with the single drug groups(both P<0.01),while the bax protein expression level in the combination group was significantly higher than that in the single drug groups(P<0.01).Conclusion Osimertinib and quercetin can exert synergistic action,which can significantly inhibit H1975 cell proliferation.The mechanism may be related to the down-regulation of p-Akt protein expression and the promotion of apoptosis.