Li Chunrui, Liu Wenli, Meng Fankai, Huang Wei, Zhou Jianfeng, Sun Hanying, Feng Yongdong
Department of Hematology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030.
J Huazhong Univ Sci Technolog Med Sci. 2003;23(3):249-53. doi: 10.1007/BF02829505.
In order to explore the molecular mechanisms of sodium butyrate and trichostatin A on K562 cell proliferation/differentiation, 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 cell surface adhesion molecules analyzed by FACS. Cell cycle distribution was studied after DNA staining by propidium iodide. Cell cycle regulatory proteins were detected by Western blot and reverse transcription-polymerase chain reaction. The results showed that sodium butyrate blocked cells mainly at the G0/G1 phase of the cell cycle, whereas trichostatin A arrested the cells at G2 phase. Sodium butyrate could down-regulate the mRNA expression of cyclin D1, but not affect its protein expression; down-regulate the protein expression of cyclin D3, but not affect its mRNA expression. Trichostatin A showed similar effects on cyclin D1 and D3 as sodium butyrate. Both sodium butyrate and trichostatin A could stimulate p21 expression of K562 cells at mRNA and protein levels. It may be concluded that sodium butyrate and trichostatin A could promote the proliferation/differentiation of the K562 cells, which might be contributed to the induced expression of cyclin D3 and p21 proteins.
为了探究丁酸钠和曲古抑菌素A对K562细胞增殖/分化的分子机制，将K562细胞分别在不存在或存在丁酸钠或曲古抑菌素A的情况下培养。通过台盼蓝拒染法测定活细胞百分比。通过硝基蓝四氮唑（NBT）还原法测定分化情况，并用荧光激活细胞分选术（FACS）分析细胞表面黏附分子。用碘化丙啶对DNA染色后研究细胞周期分布。通过蛋白质印迹法和逆转录-聚合酶链反应检测细胞周期调节蛋白。结果表明，丁酸钠主要将细胞阻滞在细胞周期的G0/G1期，而曲古抑菌素A将细胞阻滞在G2期。丁酸钠可下调细胞周期蛋白D1的mRNA表达，但不影响其蛋白表达；下调细胞周期蛋白D3的蛋白表达，但不影响其mRNA表达。曲古抑菌素A对细胞周期蛋白D1和D3的影响与丁酸钠相似。丁酸钠和曲古抑菌素A均可在mRNA和蛋白水平上刺激K562细胞的p21表达。可以得出结论，丁酸钠和曲古抑菌素A可促进K562细胞的增殖/分化，这可能与细胞周期蛋白D3和p21蛋白的诱导表达有关。
