胰岛素样生长因子1受体抑制剂NVP-AEW541对ESCC细胞增殖、迁移和侵袭的影响及其机制

Effects of insulin-like growth factor 1 receptor inhibitor NVP-AEW541 on proliferation,migration and invasion of ESCC cells

目的:探讨胰岛素样生长因子1受体(IGF-1R)抑制剂NVP-AEW541对3种食管鳞状细胞癌(ESCC)细胞KYSE30、KYSE450和TE-1增殖、迁移和侵袭的影响,初步阐明其作用机制。方法:体外培养的KYSE30、KYSE450和TE-1 ESCC细胞作为实验对象,分为对照组[0μmol·L^(-1) NVP-AEW541,1 mLRPMI 1640完全培养基+1μL二甲亚砜(DMSO)]和实验组(2、4、6、8和10μmol·L^(-1) NVP-AEW541),对照组和实验组细胞处理48 h后,CCK-8法检测各组细胞存活率;对照组和实验组(4和6μmol·L^(-1) NVP-AEW541)细胞处理0、6、12和24 h后,细胞划痕实验检测各组细胞迁移率;对照组和实验组(6μmol·L^(-1) NVP-AEW541)细胞处理48 h后,Transwell实验检测侵袭细胞数;对照组和实验组(2、4和6μmol·L^(-1) NVP-AEW541)细胞处理48 h后,Western blotting法检测各组细胞中磷酸化IGF-1R(p-IGF-1R)和上皮-间质转化(EMT)相关蛋白N-Cadherin和Snail-1的蛋白表达水平。结果:与对照组比较,实验组KYSE30、KYSE450和TE-1细胞存活率均随NVP-AEW541浓度增加而降低(P<0.05),实验组(10μmol·L^(-1))KYSE30、KYSE450和TE-1细胞存活率明显降低(P<0.01),实验组(6μmol·L^(-1))KYSE30、KYSE450和TE-1细胞迁移率随NVP-AEW541浓度增加而降低(P<0.01),实验组(6μmol·L^(-1))KYSE30、KYSE450和TE-1细胞侵袭细胞数明显减少(P<0.01),且实验组细胞中p-IGF-1R、N-Cadherin和Snail-1蛋白的表达水平均明显降低(P<0.01)。结论:NVP-AEW541能够明显抑制ESCC细胞增殖,通过抑制N-Cadherin和Snail-1蛋白表达,阻碍EMT,抑制ESCC细胞迁移和侵袭。

Objective:To explore the effects of insulin-like growth factors-1 receptor NVP-AEW541 on the proliferation,migration and invasion of three kind of esophageal squamous cell carcinoma(ESCC)cells KYSE30,KYSE450 and TE-1,and to preliminarily clarify their mechanisms.Methods:The ESCC cells KYSE30,KYSE40 and TE-1 cultured in vitro were used as experimental subjects and divided into control group(0μmol·L^(-1) NVP-AEW541,1 mL RPMI 1640 complete medium+1μL DMSO)and experimental groups(2,4,6,8 and 10μmol·L^(-1) NVP-AEW541).After the cells in control group and experimental group were treated for 48 h,CCK-8 assay was used to detect the cell survival rates in various groups;after the cells in control group and experimental groups(4 and 6μmol·L^(-1) NVP-AEW541)were treated for 0,6,12 and 24 h,cell scratch test was used to detect the cell migration rates in various groups;after the cells in control group and experimental group(6μmol·L^(-1) NVP-AEW541)were treated for 48 h,Transwell test was to used detect the number of invasion cells in various groups;after the cells in control group and experimental groups(2,4 and 6μmol·L^(-1) NVP-AEW541)were treated for 48 h,Western blotting method was used to detect the expression levels of p-IGF-1R and epithelial-mesenchymal transition(EMT)-related proteins N-Cadherin and Snail-1 proteins in the cells in various groups.Results:Compared with control group,the cell survival rates of KYSE30,KYSE450 and TE-1 cells in experimental groups were decreased with the increase of the concentration of NVP-AEW541(P<0.05);the survival rates of KYSE30,KYSE450 and TE-1 cells in experimental group(10μmol·L^(-1))were decreased(P<0.01);the cell migration rates in experiment group(6μmol·L^(-1))were decreased with the increase of NVP-AEW541 concentration(P<0.01);the number of invasion cells in experimental group(6μmol·L^(-1))was decreased(P<0.01);the expression levels of p-IGF-1R,N-Cadherin and Snail-1 proteins in the cells in experimental group were significantly decreased(P<0.01).Conclusion:NVP-AEW541 can significantly inhibit the proliferation of ESCC cells,by inhibiting the expressions of N-Cadherin and Snail-1 proteins,it prevents the EMT,and inhibit the migration and invasion of ESCC cells.