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中华肝脏外科手术学电子杂志 ›› 2016, Vol. 05 ›› Issue (04) : 265 -269. doi: 10.3877/cma.j.issn.2095-3232.2016.04.014

所属专题: 文献

基础研究

丙戊酸钠对人胆管癌细胞生长抑制作用及其机制
王兵1,(), 李红波2, 杨杰1, 邹声泉1   
  1. 1. 420100 武汉,华中科技大学同济医学院附属同济医院胆胰外科
    2. 233004 安徽省蚌埠医学院第一附属医院肿瘤外科
  • 收稿日期:2016-03-17 出版日期:2016-08-10
  • 通信作者: 王兵
  • 基金资助:
    国家科技部国际合作司项目(2010DFA31870); 国家自然基金青年项目(81502108)

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

Bing Wang1,(), Hongbo Li2, Jie Yang1, Shengquan Zou1   

  1. 1. Department of Pancreaticobiliary Surgery, Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 420100, China
    2. Department of Oncological Surgery, the First Affiliated Hospital of Bengbu Medical College, Anhui Province, Bengbu 233004, China
  • Received:2016-03-17 Published:2016-08-10
  • Corresponding author: Bing Wang
  • About author:
    Corresponding author: Wang Bing, Email:
引用本文:

王兵, 李红波, 杨杰, 邹声泉. 丙戊酸钠对人胆管癌细胞生长抑制作用及其机制[J]. 中华肝脏外科手术学电子杂志, 2016, 05(04): 265-269.

Bing Wang, Hongbo Li, Jie Yang, Shengquan Zou. Effect of sodium valproate on inhibiting the growth of human cholangiocarcinoma cells and its mechanism[J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2016, 05(04): 265-269.

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 flow 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 significantly 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 group significantly increased (LSD-t=17.557, 12.465; P<0.05). At 31 d after the first treatment, the tumor volume (338±11) mm3 in the 8.0 mmol/L experimental group significantly decresed, compared with (426±14) mm3 in the control group (t=-15.630, P<0.05).

Conclusions

Sodium valproate can significantly inhibit the growth of human cholangiocarcinoma cells, and its mechanisms include cell cycle arrest and cell apoptosis.

图1 不同浓度不同时间丙戊酸钠对TFK-1细胞的生长抑制率
图2 流式细胞术检测丙戊酸钠作用TFK-1细胞3 d的细胞凋亡率
图3 流式细胞术检测丙戊酸钠作用TFK-1细胞3 d的各周期细胞比率
图4 荧光显微镜下观察丙戊酸钠引起TFK-1细胞自噬现象
图5 丙戊酸钠实验组与对照组裸鼠胆管癌皮下种植瘤生长曲线
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