丁酸钠诱导结肠癌HT-29细胞凋亡的信号通路及对AQP3基因表达影响的研究

目的：探讨丁酸钠诱导结肠癌HT-29细胞凋亡过程的分子机理及其对水通道蛋白AQP3基因表达的影响。方法：MTT法检测不同浓度的丁酸钠对结肠癌HT-29细胞生长的影响;RT-PCR检测不同浓度的丁酸钠对细胞周期蛋白Cyclin D1、CDK4、p16和AQP3的基因表达水平,同时使用流式细胞仪检测细胞周期。结果：加入浓度超过2.5mmol/L的丁酸钠后,结肠癌HT-29细胞生长受到抑制,Cyclin D1、CDK4和AQP3的表达水平下降,p16的表达水平上调,细胞阻滞于G1/S期。结论：浓度超过2.5mmol/L的丁酸钠可以诱导结肠癌HT-29细胞凋亡,使其细胞周期阻滞于G1/S期,且这种诱导作用是剂量依赖型的,其分子通路可能是通过p16-Cyclin D1/CDK4这条信号途径,在此过程中,伴随着AQP3的基因表达水平降低。

Study on the signal pathway of cell apoptosis in HT-29 cells and the gene expression of AQP3 induced by sodium butyrate

AIM： To investigate the signal pathway of cell apoptosis in HT-29 cells and the gene expression of AQP3 induced by sodium butyrate. METHODS： MTF method was used to detect the inhibitive effect of cell growth induced by different concentrations of sodium butyrate, while RT-PCR to detect the gene expressions of Cyclin D1, CDK4, p16 and AQPS, FCM for cell cycle detection. RESULTS： With the concentration of sodium butyrate more than 2.5 mmol/L, the cell growth of HT-29 cells was inhibited, the expressions of Cyclin D1, CDK4 and AQP3 were down-regulated while the expression of p16 was up-regulated, cells were arrested in G1/S. CONCLUSION： The apoptosis of HT- 29 cells may be induced by sodium butyrate with concentrations more than 2.5 mmol/L, while the cell cycle arrested in G1/S dose-dependently. The signal pathway of cell apoptosis may be the pl6-Cyclin D1/CDK4 way, together with the down-regulation of AQV3.