癌性锚蛋白重复序列在胃癌细胞凋亡中的作用

目的 探讨癌性锚蛋白重复序列(Gankyrin)在人胃癌细胞的表达,以及在尼美舒利诱导细胞凋亡过程中的改变.方法 培养不同分化程度的人胃癌细胞系MKN28(高分化)、AGS(低分化)、MKN45(低分化)和SGC7901(中分化)],以尼美舒利处理细胞,应用四甲基偶氮唑盐试验和流式细胞术检测细胞活力及细胞凋亡,实时PCR和Western印迹法检测Gankyrin基因和蛋白表达.结果 在4种不同分化程度的人胃癌细胞系中,均存在不同水平的Gankyrin基因和蛋白表达.尼美舒利以时间-剂量依赖方式抑制AGS、FYGC7901细胞增殖.与对照组相比,尼美舒利400 μmol/L处理48 h可诱导AGS、SGC7901细胞显著凋亡(细胞凋亡率分别为0.57%±0.19%比23.30%±2.50%和0.88%±0.17%比16.80%±1.55%,P均<0.01).在AGS细胞凋亡过程中,Gankyrin基因和蛋白表达水平下降,以尼美舒利作用后24 h(0.0035±0.0014)和36 h(0.0980±0.0160)改变最为显著(对照组为0.4690±0.1190,P值均<0.01).结论 在人胃癌细胞中存在Gankyrin基因和蛋白表达.Gankyrin可能参与尼美舒利诱导的AGS胃癌细胞凋亡.

Gankyrin involved in nimesulide induced apoptosis of human gastric cancer cells

Objective To elucidate the expression of gankyrin in human gastric cancer cells and it's role in nimesulide induced apoptosis. Methods Four human gastric cancer cell lines including MKN28 (well differentiated), AGS (poorly differentiated), MKN45 (poorly differentiated), and SGC7901(moderately differentiated) were cultured and treated with nimesulide. Nimesulide induced growth inhibition and apoptosis of the cells were detected by methyl thiazolyl tetrazolium assay, and confirmed by flow cytometry. The expressions of gankyrin gene and protein were further assessed by real-time PCR and Western blotting. Results Gankyrin mRNA and protein were detected in all four human gastric cancer cell lines. The proliferations of AGS and SGC7901 cell lines were significantly suppressed by nimesulide in a time-dose dependent manner. When treated with 400 μmol/L of nimesulide for 48 hours, the significant apoptosis was found in AGS cells (23.30%±2.50%) and SGC7901 cells (16.80%±1.55% ) in comparison with controls (0.57%±0.19% and 0.88%± 0.17%, respectively, all P values <0.01). Apoptosis of AGS cells induced by nimesulide was accompanied by a considerably decreased gankyrin expression that was more significant at 24 hours (0.0035±0.0014) and 36 hours (0.0980±0.0160) in comparison with controls (0.4690±0.1190, all P values<0.01). Conclusion Gankyrin expresses in human gastric cancer cell lines and may be involved in nimesulide induced apoptosis of AGS cells.