摘要
目的：观察丁酸钠（SB）对人喉癌Hep-2细胞的生长抑制作用，探讨其对细胞凋亡及细胞周期的影响。方法：以MTT法检测0．625～10．000mmol／LSB对Hep-2细胞生长的抑制作用；以TUNEL法检测1．250～5.000mmol／LSB作用24h、48h、72h后的细胞凋亡情况；以琼脂糖凝胶电泳法检测2．500mmol／LSB作用24h、48h、72 h后的细胞DNA片段；应用流式细胞仪分析1．250mmol／L、2．500mmol／L、5．000mmol／LSB作用24h后细胞凋亡率和细胞周期变化；采用免疫组织化学sP法检测2．500mmol／LSB作用24h、48h、72h后细胞抗凋亡蛋白Survivin表达的变化。结果：SB对Hep-2细胞的生长抑制作用具有时间和剂量关系（不同时间组、不同剂量组间比较，P均〈0．01）；细胞凋亡指数随sB剂量升高、作用时间延长而增加（P均〈0．01）；琼脂糖凝胶电泳可见特征性的凋亡细胞DNA梯状条带；流式细胞术显示细胞凋亡率随SB剂量升高而增加（P〈0．01），细胞周期阻滞于G0／G1期；2．500mmol／LSB作用72h，Survivin表达阳性细胞的光密度值由作用前的0．164±0．009下降至0．045±0．006（P〈0．01）。结论：sB能有效抑制Hep-2细胞的生长，其机制可能与抑制细胞Survivin蛋白表达进而诱导细胞凋亡以及参与细胞周期调控有关。
Aim ： To study the effects of sodium butyrate（SB） on cell growth, apoptosis, and cell cycle in Hep-2 cells. Methods ：The proliferative activity of Hep-2 cells treated with SB from 0. 625 mmol/L to 10. 000 mmol/L was assessed by MTT assay. Cell apoptosis in the cells treated with SB from 1. 250 mmol/L to 5. 000 mmol/L was detected by TUNEL method. DNA fragmentation in the cells treated with SB at 2. 500 mmol/L was examined by agarose gel electrophoresis. The changes of cell apoptosis rate and cell cycle distribution were analyzed by flow cytometry. Expression of Survivin protein in the cells treated with SB at 2. 500 mmol/L was detected by immunohistochemistry SP method. Results ： A time- and dose-dependent inhibition was detected in Hep-2 cells treated with SB （P 〈0.01 ）. Apoptosis index（AI） in the cells treated with SB increased with the increased concentration and prolonged action time （P 〈0.01 ）. Characteristic DNA ladders could be demonstrated in agarose gel by electrophoresis. Flow cytometry revealed that cell apoptosis rate increased with the increased concentration of SB （P 〈 0.01 ） , and the cells were blocked in G0/G1 phase. After the treatment of SB at 2.500 mmoL/L for 72 h,＂ the optical density of Survivin protein in the cells decreased to 0. 045 ± 0. 006, which was 0. 164 ± 0. 009 before the treatment （P 〈0.01 ）. Conclusion： SB could inhibit growth of Hep-2 cells effectively. Its mechanism may be related to inhibiting expression of Survivin protein and further inducing cell apoptosis, and taking part in regulating cell cycle.
出处
《郑州大学学报（医学版）》
CAS
北大核心
2007年第2期318-322,共5页
Journal of Zhengzhou University(Medical Sciences)
