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目的探讨激发CD40信号对胃癌细胞生长、增殖的影响以及胃癌细胞株AGS基因表达谱的变化。方法四甲基偶氮唑盐(MTT)微量酶反应比色法检测不同浓度可溶性CD40配体(sCD40L)作用后胃癌细胞株AGS的存活率,以选择最佳干预浓度。流式细胞术检测sCD40L最佳干预浓度作用后胃癌细胞株AGS细胞周期和凋亡的变化。以sCD40L最佳干预浓度与胃癌细胞株AGS共育48h,基因表达谱芯片Affymetrix U133A 2.0检测胃癌细胞株AGS基因表达谱变化。结果sCD40L的最佳干预浓度为2μg/ml;与胃癌细胞株AGS共育48h后,sCD40L可使AGS细胞株生长抑制,细胞周期阻滞在G1期。激发CD40信号前、后,胃癌细胞株AGS差异表达基因414个,其中上调基因209个,下调基因205个。表达水平显著改变的基因共45个(P<0.01),其中表达上调基因38个,下调基因7个。结论sCD40L对高表达CD40的胃癌细胞有明显的生长抑制作用。AGS细胞差异基因功能涉及代谢、凋亡及信号转导等,且可能主要活化p38丝裂原活化蛋白激酶和应激活化蛋白激酶/c-Jun氨基末端激酶信号通路,提示CD40信号对CD40阳性表达胃癌细胞的生物学行为可能具有重要的调节作用。
Objective To investigate the effect of CD40 signaling on the growth and proliferation of gastric cancer cells and the changes of AGS gene expression in gastric cancer cell lines. Methods MTT assay was used to determine the survival rate of gastric cancer cell line AGS after different concentrations of soluble CD40 ligand (sCD40L) were assayed by MTT colorimetric assay to select the best intervention concentration. Flow cytometry was used to detect the changes of cell cycle and apoptosis in gastric cancer cell line AGS treated with sCD40L at the best concentration. The optimal concentration of sCD40L was co-cultured with gastric cancer cell line AGS for 48h, and the gene expression profile Affymetrix U133A 2.0 was used to detect the gene expression profiles of AGS in gastric cancer cell lines. Results The optimal concentration of sCD40L was 2μg / ml. After co-cultured with gastric cancer cell line AGS for 48h, sCD40L inhibited the growth of AGS cell line and inhibited the cell cycle arrest in G1 phase. There were 414 differentially expressed genes in gastric cancer cell line AGS before and after the stimulation of CD40 signal, including 209 up-regulated genes and 205 down-regulated genes. There were 45 genes with significant changes (P <0.01), including 38 up-regulated genes and 7 down-regulated genes. Conclusion sCD40L has obvious growth inhibitory effect on gastric cancer cells with high expression of CD40. AGS cell differential gene function involves metabolism, apoptosis and signal transduction, and may mainly activate p38 mitogen-activated protein kinase and stress activated protein kinase / c-Jun N-terminal kinase signaling pathway, suggesting CD40 signal on CD40 positive expression The biological behavior of gastric cancer cells may have an important regulatory role.