IGF-1R抑制剂NVP-AEW541对卵巢癌移植瘤中休眠细胞的作用

Effect of IGF-1R inhibitor NVP-AEW541 on the dormant cells isolated from ovarian cancer xenografts

目的 :探讨胰岛素样生长因子1受体(insulin-like growth factor-1 receptor,IGF-1R)抑制剂NVP-AEW541对卵巢癌移植瘤中休眠的SKOV3-PKH26^(hi)细胞增殖和凋亡的影响,以及其可能的作用机制。方法:应用CCK-8法检测不同浓度的NVP-AEW541(1、5、10和15μmol/L)对SKOV3-PKH26^(hi)细胞增殖的影响。NVP-AEW541处理SKOV3-PKH26^(hi)细胞48 h后,应用FCM法检测细胞周期及细胞凋亡。蛋白质印迹法检测NVP-AEW541对SKOV3-PKH26^(hi)细胞中细胞周期蛋白D1(cyclin D1)、caspase-3、Bcl-2、Bax、磷酸化蛋白激酶B(phosphoprotein kinase B,p-PKB,又称p-Akt)、磷酸化IGF-1R(phospho-IGF-1R,p-IGF-1R)及磷酸化糖原合成酶激酶-3β(phospho-glycogen synthase kinase-3β,p-GSK-3β)蛋白表达的影响。结果 :NVP-AEW541可抑制SKOV3-PKH26^(hi)细胞增殖,呈剂量和时间依赖性(P值均<0.01)。NVP-AEW541将SKOV3-PKH26^(hi)细胞阻滞于G_0/G_1期,并促进细胞早期凋亡和晚期凋亡(P值均<0.01)。与对照组(SKOV3-PKH26^(hi)细胞未进行任何处理)比较,NVP-AEW541明显下调SKOV3-PKH26^(hi)细胞中cyclin D1和Bcl-2蛋白表达水平(P值均<0.05),明显上调caspase-3和Bax蛋白表达水平(P值均<0.05),p-IGF-1R、p-Akt和p-GSK-3β的表达水平也明显下调(P值均<0.01)。结论 :IGF-1R抑制剂NVP-AEW541可以通过下调Akt的磷酸化水平,抑制SKOV3-PKH26^(hi)细胞增殖,并促进细胞凋亡。

Objective： To investigate the effects of NVP-AEW541, an inhibitor of insulin-like growth factor 1 receptor（IGF-1R）, on the proliferation and apoptosis of dormant SKOV3-PKH26^（hi） cells isolated from ovarian cancer xenografts, and to explore the possible mechanism.Methods： The effect of NVP-AEW541（1, 5, 10, 15 μmol/L） on the proliferation of SKOV3-PKH26^（hi） cells was detected by CCK-8 assay. After the SKOV3-PKH26^（hi） cells were treated with NVP-AEW541 for 48 h, FCM was performed to detect the cell cycle and apoptosis. The effects of NVP-AEW541 on the expressions of cyclin D1, caspase-3, Bax, Bcl-2, phospho-protein kinase B（p-PKB, also known as p-Akt）, phospho-IGF-1R（p-IGF-1R） and phospho-glycogen synthase kinase-3β（p-GSK-3β） in SKOV3-PKH26^（hi） cells were detected by Western blotting.Results： NVP-AEW541 had anti-proliferative effect on the SKOV3-PKH26^（hi） cells in a dose-and time-dependent manner（both P〈 0.01）. NVP-AEW541 arrested the cell cycle at G_0/G_1 phase, and promoted the early and late apoptosis（all P〈 0.01）. The expressions of cyclin D1 and Bcl-2 proteins were decreased（both P〈 0.05）, and the expressions of caspase-3 and Bax proteins were increased（both P〈 0.05） as compared with the control group（SKOV3-PKH26^（hi） cells without any treatment）. NVP-AEW541 downregulated significantly the expression levels of p-IGF-1R, p-Akt and p-GSK-3β proteins（all P〈 0.01）.Conclusion： IGF-1R inhibitor NVP-AEW541 is effective to inhibit the proliferation of SKOV3-PKH26^（hi） cells and to promote apoptosis by reducing the phosphorylation level of Akt.