丙戊酸钠对人胆管癌细胞生长抑制作用及其机制

Effect of sodium valproate on inhibiting the growth of human cholangiocarcinoma cells and its mechanism

目的探讨丙戊酸钠对人胆管癌细胞生长抑制作用及其机制。方法分别以0.5、1.0、2.0、4.0、8.0 mmol/L浓度的丙戊酸钠作用于人胆管癌TFK-1细胞1、3、5 d,另设对照组和空白组。采用CCK-8法检测丙戊酸钠对人胆管癌细胞生长抑制作用,流式细胞术检测丙戊酸钠对细胞凋亡率及细胞周期的影响。采用裸鼠TFK-1细胞皮下种植瘤模型,设立实验组和对照组,观察丙戊酸钠对瘤体生长的影响。两组实验数据比较采用t检验,多组比较采用单因素方差分析和LSD-t检验。结果丙戊酸钠对人胆管癌TFK-1细胞的生长抑制作用呈浓度和时间依赖性。随着药物浓度的增加,细胞凋亡率和G2/M期细胞比率明显增加。8.0 mmol/L实验组3 d时细胞凋亡率(57.5±6.2)%较对照组的(8.6±2.3)%明显增加,G2/M期细胞比率(42.5±5.4)%较对照组的(12.0±2.6)%明显增加(LSD-t=17.557,12.465;P<0.05)。第一次治疗后31 d,实验组肿瘤体积(338±11)mm3较对照组的(426±14)mm3明显减少(t=-15.630,P<0.05)。结论丙戊酸钠对人胆管癌细胞的生长有明显的抑制作用,其机制包括细胞周期阻滞和细胞凋亡。

Objective To investigate the effect and mechanism of sodium valproate on inhibiting the growth of human cholangiocarcinoma cells.Methods Human cholangiocarcinoma TFK-1 cells were respectively treated with 0.5,1.0,2.0,4.0 and 8.0 mmol/L of sodium valproate for 1,3 and 5 d.And the control and blank groups were also established.The inhibitory effect of sodium valproate on the growth of human cholangiocarcinoma cells was detected by cell counting kit（CCK）-8 assay.The effect of sodium valproate on the cell apoptosis rate and cell cycle was detected by fl ow cytometry.The nude mice subcutaneous implantation tumor models of TFK-1 cells was established and the experimental and control groups were assigned to observe the effect of sodium valproate on the growth of tumor.The experiment data in two groups were compared using t test,and the multi-group comparison was conducted using one way analysis of variance and LSD-t test.Results The inhibitory effect of sodium valproate on the growth of human cholangiocarcinoma TFK-1 cells was observed in a dose-and time-dependent manner.As the concentration of sodium valproate increased,the cell apoptosis rate and the percentage of cells in the G2/M phase significantly increased.Compared with（8.6±2.3）% in the control group,the cell apoptosis rate（57.5±6.2）% in the 8.0 mmol/L experimental group signifi cantly increased at 3 d.Compared with（12.0±2.6）% in the control group,the percentage of cells in the G2/M phase（42.5±5.4）% in the 8.0 mmol/L experimental groupsignifi cantly increased（LSD-t=17.557,12.465;P〈0.05）.At 31 d after the fi rst treatment,the tumor volume（338±11） mm3 in the 8.0 mmol/L experimental group signifi cantly decresed,compared with（426±14） mm3 in the control group（t=-15.630,P〈0.05）.Conclusions Sodium valproate can signifi cantly inhibit the growth of human cholangiocarcinoma cells,and its mechanisms include cell cycle arrest and cell apoptosis.