硒代甲硫氨酸联合5-氟尿嘧啶对人胃癌MKN-45细胞的协同抑制作用

The synergistic inhibition effect of selenomethionine combined with 5-l uorouracil on human gastric cancer MKN-45 cells

目的 :研究硒代甲硫氨酸(selenomethionine,Se Met)联合5-氟尿嘧啶(5-l uorouracil,5-FU)对人胃癌MKN-45细胞的抑制作用,并初步探讨其可能的作用机制。方法 :采用不同浓度的Se Met与5-FU单独或联合作用于MKN-45细胞。细胞计数试剂盒-8(cell counting kit-8,CCK-8)法检测细胞增殖的抑制率,并采用Chou-Talalay联合指数法分析两药之间的相互作用;FCM法检测细胞的凋亡率;细胞划痕实验检测细胞的迁移能力;Transwell小室侵袭实验检测细胞的侵袭能力;蛋白质印迹法检测细胞中p53、p21、p27及细胞周期蛋白cyclin D1的表达水平。结果 :不同浓度的Se Met与5-FU单药或联合干预后,MKN-45细胞的增殖均明显受到抑制(P值均<0.01),并呈剂量依赖性,两药有协同作用。Se Met与5-FU联合用药组较空白对照组和各单药组细胞的凋亡率增加,迁移及侵袭能力降低(P值均<0.01)。联合用药组的p53、p21和p27蛋白的表达水平均较空白对照组和各单药组明显上调(P值均<0.01),而cyclin D1的表达水平明显下调(P值均<0.01)。结论 :Se Me联合5-FU能协同抑制人胃癌MKN-45细胞的增殖、迁移及侵袭,促进凋亡,其机制可能与调节MKN-45细胞中p53、p21、p27及cyclin D1蛋白的表达水平有关。

Objective：To investigate the inhibition effect of selenomethionine（SeMet） combined with 5-fluorouracil（5-FU） on human gastric cancer MKN-45 cells,and to explore the possible mechanism.Methods：The MKN-45 cells were treated with different concentrations of SeMet or 5-FU alone or in combination.The proliferation inhibitory rate of MKN-45 cells was detected by cell counting kit-8（CCK-8） method.The interaction of SeMet and 5-FU was estimated by Chou-Talalay combination index method.The apoptosis and the migration and invasion abilities of MKN-45 cells were detected by flow cytometry,scratch-wound assay and Transwell chamber assay,respectively.The expression levels of p53,p21,p27 and cyclin Dl proteins in MKN-45 cells were examined by Western blotting.Results：The proliferation inhibitory rates of MKN-45 cells in single drug groups（SeMet or5-FU） and the combination group（SeMet and 5-FU） were increased as compared with that in the control group（without any treatment）（P 0.01） in a dose-dependent manner.SeMet and 5-FU had a synergistic effect.As compared with the control group and the single drug groups,the apoptosis rate of MKN-45 cells after treatment with combination of SeMet and5-FU was increased and the migration and invasion abilities were decreased（all P 0.01）;the expression levels of p53,p21 and p27 proteins in combination group were significantly up-regulated（all P 0.01）,whereas the expression of cyclin Dl protein was down-regulated（both P 0.01）.Conclusion：SeMet in combination with 5-FU can synergistically inhibit the proliferation,migration and invasion of MKN-45 cells as well as promote apoptosis.This mechanism may be related to regulating the expressions of p53,p21,p27 and cyclin Dl proteins in MKN-45 cells.