体外转染Kiss-1基因对人黑色素瘤细胞A375生长增殖的影响

Effect of anti-oncogene Kiss-1 gene on cell proliferation in human melanoma cells

目的 探讨转染Kiss-1基因对人黑色素瘤细胞A375生长增殖的影响,筛选出有意义的肿瘤治疗的生物学靶标.方法 在人黑色素瘤细胞A375中转染Kiss-1基因,经G418筛选,建立稳定高表达Kiss-1蛋白的细胞系.Western blot方法 检测Kiss-1蛋白以证实转染成功.流式细胞术检测Kiss-1基因对A375生长增殖的影响.结果 稳定转染Kiss-1基因的A375细胞中不仅Kiss-1蛋白稳定表达（1.20±0.21）高于对照组（0.60±0.41）,Kiss-1基因转染A375细胞48 h后对其具有凋亡的作用（凋亡率为28.42%）高于对照组（2.12%）.结论 外源性Kiss-1基因导入A375细胞后,可诱导A375细胞其凋亡,从而抑制肿瘤细胞增殖.

Objective To investigate the effect of Kiss-1 on cell proliferation in human melanoma cancer cell,and screen a target for tumor biotherapy. Methods The eukaryotic expression vectors with Kiss-1 gene were used to transfect A375 cells by liposome-mediated transfection in vitro. The cells were selected by G418. The expression of Kiss-1 gene was confirmed by Western Blotting. FACS was used to detect the ability of cells proliferation and apoptosis. Results There were significantly high expression of Kiss-1 protein in the transfected A375 cells（1.20 ±0.21 vs control 0.60 ＋ 0. 41）, whose growth rate was significantly slower than that of control. Flow cytometry showed that apoptosis of A375 cells transfected Kiss-1 were induced（7.23% vs control 4.62%）. Conclusion When transfected into A375 cells,exogenous Kiss-1 could cause the cell growth arrest to G1 ,and induce cells apoptosis.