阿维A酸诱导皮肤鳞癌细胞凋亡及对caspases蛋白的影响

目的 探讨阿维A酸对人皮肤鳞癌细胞系SCC13细胞的生长抑制、凋亡的诱导作用以及对caspases蛋白的影响.方法 以体外培养的人皮肤鳞癌细胞系SCC13细胞为研究对象,采用MTT法观察5种浓度(5×10-7、1×10-6、5×10-6、1×10-5、5×10-5moL/L)阿维A酸对SCC13细胞的生长抑制作用.ELISA检测凋亡的诱导情况.流式细胞仪检测细胞周期.Western blot分析caspase-8和caspase-9的蛋白表达.结果 阿维A酸能显著抑制人皮肤鳞癌细胞的生长(第1、3、5天F值分别为83.64、96.34和123.17)、诱导其凋亡(第1、3、5天F值分别为74.45、107.37和64.28),具有浓度及时间依赖性(P均<0.05).阿维A酸能将细胞周期阻滞于G1期,作用SCC13细胞72 h后,G1期细胞比率明显上升为77.05%,而对照组仅为63.55%.阿维A酸作用SCC13细胞12 h后开始活化caspase-8蛋白,24 h后开始活化caspase-9蛋白,并具有时间依赖性(P均<0.01).结论 阿维A酸能抑制皮肤鳞癌细胞生长并诱导凋亡,这一效应可能由Fas死亡受体和线粒体途径共同介导.

Apoptosis of acitretin-induced cutaneous squamous cell carcinoma cells and effects on cuspases expressions.

Objective To investigate the inducing effect of acitrotin on the growth and apoptosis of human cutaneous squamous cell carcinoma cell line SCC13 and on caspases expression.Methods Human cutaneous squa-mous cell carcinoma cell line SCC13 was treated with five different concentrations of acitrefin 5×10-7,1×10-6,5×10-6,1×10-5,5×10-5mol/L].Cell proliferation was evaluated by MTT assay.Apoptosis was assessed by en-zyme-linked immunosorbent assay.The cell cycle was assessed by flow cytometry.The protein expressions of caspase-8 and caspase-9 were examined with Western blot.Results Acitretin inhibited the growth ( F = 83.64,96.34 and 123.17, respectively on the first, third and fifth day)and induced the apoptosis of SCC13 cells(F=74.45,107.37,and 64.28, respectively on the first, third and fifth day) in a dose- and time-dependent manner(P<0.05).Acitretin altered cell cycle distribution of SCC13 cells as compared with controls, the G1-phase population increased by 77.66% 72 hours after acitretin treatment, while the control increased only by 63.55%.An active fragment of caspase-8 occurred following 12 hours treatment of acitretin on SCC13 cells, whereas the caspase-9 active fragment occurred 24 hours after acitretin treatment, which increased time-dependently (P<0.01).Conclusion Acitretin plays an important role on the growth inhibition and apoptosis of cutaneous squamous cell carcinoma cells, which may be affected through both Fas receptor way and mitochondria way.