苯丁酸钠对人舌鳞状细胞癌细胞凋亡影响机制的探讨

摘要

目的 探讨苯丁酸钠对人舌鳞状细胞癌Tca8113和TCSSA细胞株的生长、凋亡的影响及其分子机制,为临床治疗提供理论依据.方法 应用甲基噻唑基四唑(MTT)法检测苯丁酸钠对人舌鳞状细胞癌细胞株的抑制作用,采用流式细胞仪研究苯丁酸钠对人舌鳞状细胞癌细胞株的细胞周期阻滞和诱导凋亡作用,应用蛋白质印迹法和反转录聚合酶链反应(RT-PCR)检测p21和生存素基因的转录和表达.结果 苯丁酸钠对两种细胞株增殖均有明显的抑制作用;流式细胞仪检测显示苯丁酸钠诱导细胞株G1期阻滞,碘化丙啶-异硫氰酸荧光素标记的磷脂酰结合蛋白双标结果表明,苯丁酸钠能诱导细胞株凋亡,使p21的mRNA及蛋白质表达升高,与对照组比较,Tca8113细胞株的p21 mRNA及蛋白质分别增加0.09±0.08和0.72±0.10,TCSSA细胞株的p21mRNA及蛋白质分别增加1.34 4±0.12和1.56±0.09(P<0.05).生存索的mRNA及蛋白质表达下降.与对照组比较,Tca8113细胞株的生存素mRNA及蛋白质分别降低1.10±0.05和1.14±1.10,TCSSA细胞株的生存素mRNA及蛋白质分别降低1.02±0.08和0.94±0.09(P<0.05).苯丁酸钠诱导p21表达上升和生存素表达下降,二者的mRNA水平变化呈显著负相关(rs=-0.532,P<0.001),蛋白表达变化同样呈显著负相关(rs=-0.564,P<0.001).结论 苯丁酸钠能抑制人舌鳞状细胞癌细胞株增殖,诱导细胞G1期阻滞和细胞凋亡.%Objective To examine the effects of sodium phenylbutyrate on the apoptosis of human tongue squamous cancer cell line and expression of p21 and survivin genes.Methods The inhibition effects of sodium phenylbutyrate on Tca8113 and human tongue squamous cell carcinoma (TCSSA) cell lines were detected by methyl thiazoly terazolium (MTT) and the apoptosis of the cancer cells after being induced by sodium phenylbutyrate examined by flow cytometry(FCM).The expression of p21 and survivin genes were observed with Western blotting and RT-PCR Results Compared with control group,the level of p21 mRNA and protein of Tca8113 cellline increased to 0.09±0.08 and increased 0.72±0.10,that of TCSSA cellline increased 1.34±0.12 and 1.56±0.09(P <0.05).Compared with control group,the level of surrive mRNA and protein of Tca8113 cellline decreased to 1.10±0.05 and 1.14±1.10,that of TCSSA cellline decreased to 0.12±0.08 and 0.94±0.09 (P < 0.05) .Sodium phenylbutyrate inhibited the cell proliferation ,promoted cell apoptosis and arrested the cells in G1/G0 phase.The amount of p21 mRNA and protein were increased,and the expression of survivin gene was decreased.Conclusions Sodium phenylbutyrate exhibited remarkable inhibitory effects on human tongue squamous cancer cell proliferation and induced cancer cell apoptosis.The mechanism may be due to up-regulation of p21 gene and downregulation of survivin gene.The mRNA level of p21 gene and survivin gene showed a strong correlation.