丁酸钠对人胰腺癌细胞ASPC-1生长的影响及其机制的研究

目的 探讨丁酸钠对人胰腺癌ASPC-1细胞生长的影响并探讨其作用机制.方法 使用不同浓度丁酸钠处理ASPC-1细胞.MTT法观察肿瘤细胞的增殖;流式细胞仪检测细胞凋亡和细胞周期的变化;Western印迹法检测细胞内p21、p53、bcl-2、bax蛋白表达的变化,实时定量PCR检测p21和bcl-2的表达.结果 丁酸钠能明显抑制ASPC-1细胞的增殖,其作用具有明显的时间和剂量依赖性.丁酸钠处理24 h后ASPC-1细胞凋亡率明显提高(P＜0.05),S期细胞比例显著降低(P＜0.05),G0/G1期细胞比例显著升高(P＜0.05).p21、bax蛋白表达明显上调(P＜0.05);bcl-2蛋白的表达显著降低(P＜0.05);p53蛋白表达无明显变化(P＞0.05).结论 丁酸钠可以通过诱导胰腺癌细胞的凋亡和周期阻滞而抑制癌细胞生长;其发生机制可能与下调抗凋亡基因bcl-2、上调促凋亡基因bax和肿瘤抑制基因p21的表达有关.

Abstract：

Objective To investigate the effects of sodium butyrate(NaBT) on proliferation of human pancreatic cancer cell line ASPC-1 and explore the possible mechanism. Methods The methylthiazolyl tetrazolium assay (MTT) method was used to detect cell proliferation and draw a curve. The cell apoptosis and cell cycle were determined with flow cytometry. Western blot was used to study the effect NaBT on the pancreatic cancer cells and explore its mechansim. Real-time PCR was employed to assess the expression levels of p53, p21, bcl-2 and cell cycle regulation gene p21. Results After incubation with different concentrations of NaBT for 24 to 72 h, ASPC-1 cell proliferation was inhibited dramatically. NaBT induced an increase of G0/G1 phase cells and a significant decrease in the ratio of S phase cells. The expression of p21 and bax was up-regulated at protein and mRNA level. The expression of bcl-2 was down-regulated at protein and mRNA level. There was no significant difference in the expression of p53 at protein and mRNA level. Conclusion TSA-induced growth inhibition is associated with a block in the G0/G1 phase and apoptosis, which may occur through down-regulating the expression of apoptosis gene bcl-2 and up-regulating the expression of cell cycle regulation gene p21and pro-apoptotic gene bax.

Effects of sodium butyrate on proliferation and differentiation of human pancreatic cancer cell line ASPC-1

Objective To investigate the effects of sodium butyrate(NaBT) on proliferation of human pancreatic cancer cell line ASPC-1 and explore the possible mechanism. Methods The methylthiazolyl tetrazolium assay (MTT) method was used to detect cell proliferation and draw a curve. The cell apoptosis and cell cycle were determined with flow cytometry. Western blot was used to study the effect NaBT on the pancreatic cancer cells and explore its mechansim. Real-time PCR was employed to assess the expression levels of p53, p21, bcl-2 and cell cycle regulation gene p21. Results After incubation with different concentrations of NaBT for 24 to 72 h, ASPC-1 cell proliferation was inhibited dramatically. NaBT induced an increase of G0/G1 phase cells and a significant decrease in the ratio of S phase cells. The expression of p21 and bax was up-regulated at protein and mRNA level. The expression of bcl-2 was down-regulated at protein and mRNA level. There was no significant difference in the expression of p53 at protein and mRNA level. Conclusion TSA-induced growth inhibition is associated with a block in the G0/G1 phase and apoptosis, which may occur through down-regulating the expression of apoptosis gene bcl-2 and up-regulating the expression of cell cycle regulation gene p21and pro-apoptotic gene bax.