丁酸钠和曲古抑菌素A促进K562细胞分化机制的研究及其对比

The effect and comparison of sodium butyrate and trichostatin A on the differentiation of K562

目的 探讨丁酸钠与曲古抑菌素A(TSA)促进K5 6 2细胞分化的机制 ,并对比二者的异同。方法 台盼蓝拒染实验观察药物对细胞生长曲线的影响 ;四氮唑盐还原试验和细胞表面分化抗原检测观察药物对细胞的分化作用 ;流式细胞仪分析细胞周期 ;RT PCR和WesternBlot分别检察细胞周期蛋白在mRNA和蛋白质水平的表达。结果 丁酸钠和TSA分别将细胞主要阻滞于G0 /G1期和G2 期 ;丁酸钠下调cyclinD1mRNA的水平基本上不影响其蛋白质的表达 ,上调cyclinD3蛋白质表达水平基本上不影响其mRNA的水平 ;TSA与丁酸钠的作用相同 ;二者均可以在mRNA和蛋白质水平刺激K5 6 2细胞 p2 1的表达。 结论 丁酸钠和TSA促进K5 6 2的分化是通过诱导 p2 1和cyclinD3蛋白质的表达而完成的 ,丁酸钠和TSA均有望成为治疗慢性髓系白血病的药物。

AIM To analyse the mechanisms of sodium butyrate action on K562 cell differentiation and to compare them with those of trichostatin A. METHODS K562 cells were grown in the absence or presence of sodium butyrate or trichostatin A. The percentage of viable cells was determined by trypan blue exclusion. Differentiation was determined by nitro blue tetrazolium (NBT) reduction and analysed cell surface adhesion molecules by FACS. Cell cycle distribution was studied after DNA staining by propidium iodide. Cell cycle regulatory proteins were studied by Western Blot and reverse transcription polymerase chain reaction. RESULTS Sodiun butyrate blocked cells mainly in the G 0/G 1 phase of the cell cycle, whereas trichostatin A was inhibitory in G 2 phases. Sodium butyrate inhibited the mRNA expression of cyclinD1 without affecting its protein expression and stimulated the protein expression of cyclinD3 without affecting its mRNA expression. Trichostatin A showed similar effects on cyclinD1 and D3. Sodium butyrate and trichostatin A stimulated p21 expression both at the mRNA and protein levels. CONCLUSION The sodium butyrate and trichostatin A effect on cell differentiation may be linked to its ability to induce expression of cyclinD3 and p21 protein. Our observations support the claim for the therapeutic potential of sodium butyrate and trichostatin A in the treatment of chronic myelocytic leukemia.