摘要
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.
作者
周楠
回丽敏
刘振兴
王利飞
ZHOU Nan, HUI Limin, LIU Zhenxing, WANG Lifei(Department of Gynecology, Xingtai People's Hospital, Xingtai 054001, Hebei Province, China)
出处
《肿瘤》
CAS
CSCD
北大核心
2018年第10期942-950,共9页
Tumor
基金
邢台市科技计划项目(编号:2016ZC292)~~