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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 study the inhibitory effect of selenomethionine (Se Met) combined with 5-fluorouracil (5-FU) on human gastric cancer MKN-45 cells and to explore its possible mechanism. METHODS: MKN-45 cells were treated with different concentration of Se Met and 5-FU alone or in combination. Cell counting kit-8 (CCK-8) was used to detect the inhibition rate of cell proliferation, and the Chou-Talalay joint index method was used to analyze the interaction between the two drugs; FCM was used to detect the apoptosis rate Cell migration assay was performed by cell scratch assay. Transwell invasion assay was used to detect cell invasion. Western blotting was used to detect the expression of p53, p21, p27 and cyclin D1. Results: Proliferation of MKN-45 cells was significantly inhibited by different concentrations of Se Met and 5-FU (all P <0.01), in a dose-dependent manner. The two drugs had synergistic effects. Compared with the blank control group and the single drug group, the apoptosis rate of Se Met and 5-FU combination group increased, while the migration and invasion ability decreased (P <0.01). The expression levels of p53, p21 and p27 protein in the combination group were significantly up-regulated compared with the blank control group and each single drug group (P <0.01), while the expression of cyclin D1 was significantly decreased (P <0.01). CONCLUSION: Se Me combined with 5-FU can synergistically inhibit the proliferation, migration and invasion of human gastric cancer MKN-45 cells and promote apoptosis. The mechanism may be related to the regulation of the expression of p53, p21, p27 and cyclin D1 in MKN-45 cells related.