摘要
目的探讨钙黏蛋白E、N(E-,N-cadherin)在依维莫司(RAD001)抑制胃癌SGC7901细胞迁移中的作用机制,及其在判断胃癌患者预后方面的作用。方法用不同终浓度的RAD001干预胃癌SGC7901细胞,用Transwell小室迁移实验检测细胞迁移能力,用免疫细胞化学法检测E-,N-cadherin的表达,用Western Blot检测不同药物浓度E-,N-cadherin的变化。在进一步探讨西罗莫司靶蛋白(mTOR)下游对E-,N-cadherin的作用中,用Western Blot检测mTOR下游对E-,N-cadherin蛋白表达的效果。结果当药物浓度达到1.25 nmol·L^(-1)及以上时,细胞迁移能力随药物剂量的增加逐渐降低;浓度达到2.5 nmol·L^(-1)及以上时,细胞中E-cadherin的蛋白表达明显增加。4EBP1-siRNA组的E-cadherin蛋白表达水平下降为0.48±0.08,而N-cadherin蛋白表达水平上升达1.35±0.12,与其余3组相比,差异均有统计学意义(均P<0.05);p70S6K-siRNA转染组的E-,N-cadherin蛋白表达水平分别为0.64±0.06和1.08±0.96,与其余3组相比,差异均无统计学意义(均P>0.05)。E-cadherin在胃癌患者中低表达,而N-cadherin在胃癌患者中高表达。E-cadherin的低表达和N-cadherin的高表达与胃癌的预后不良相关。结论 RAD001可能是通过抑制mTOR/重组人翻译起始因子4E结合蛋白1(4EBP1)/cadherin的通路而发挥作用,并且E-cadherin可以作为判断患者预后的潜在指标之一。
Objective To investigate the specific mechanism of E-,N-cadherin in evenlimus(RAD001)inhibiting the migration of gastric cancer SGC7901 cells and their role in predicting the prognosis of gastric cancer patients.Methods Gastric cancer SGC7901 cells were treated with different final concentrations of RAD001.The migration ability of SGC7901 cells was detected by Transwell cell migration assay,the expressions of E-,N-cadherin were detected by immunocytochemistry,and the changes of E-,N-cadherin were detected by Western Blot.The effect of mammalian target of rapamycin(mTOR)downstream on E-,N-cadherin protein expression was also detected by Western Blot.Results When the concentration of RAD001 reached 1.25 nmol·L^(-1) or above,the cell migration ability decreased with the increase of drug dosage;when the concentration reached 2.5 nmol·L^(-1) or above,the protein expression of E-cadherin increased significantly.The protein expression level of E-cadherin in 4 EBP1-siRNA group decreased to 0.48±0.08,while that of N-cadherin increased to 1.35±0.12,and the differences were statistically significant compared with other three groups(all P<0.05);the expression levels of E-,N-cadherin in p70 S6 K-siRNA transfection group were 0.64±0.06 and 1.08±0.96,there were no significant differences compared with other three groups(all P>0.05).The low expression of E-cadherin and high expression of N-cadherin were associated with poor prognosis of gastric cancer.Conclusion RAD001 may play a role by inhibiting the mTOR/recombinant human eukaryotic translation initiation factor 4 E-binding protein 1/cadherin pathway,and E-cadherin may be one of the prognostic indicators of patients.
作者
陆晨晨
胡风兰
刘清华
刘莹
LU Chen-chen;HU Feng-lan;LIU Qing-hua;LIU Ying(Teaching and Research Section of Pathology,Xuzhou Medical University,Xuzhou 221004,Jiangsu Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2021年第7期837-841,共5页
The Chinese Journal of Clinical Pharmacology
基金
江苏省卫生厅指导性科研课题资助项目(Z201016)。
