丁酸钠联合全反式维甲酸对MDS细胞株SKM-1的诱导分化作用及其机制初步探讨

Effect of Sodium Butyrate in Combination with ATRA on the Proliferation/Differen tiation of MDS cell line SKM-1

本研究探讨丁酸钠 (NaB)抑制MDS细胞株SKM 1细胞生长、诱导其分化的分子机制 ,并研究其与全反式维甲酸 (ATRA)的协同作用。用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 ;流式细胞术分析细胞周期 ;RT PCR检察D型细胞周期蛋白、CDK和P2 1在mRNA水平的表达。结果表明 :NaB和 (或 )ATRA均可抑制SKM 1细胞的生长 ,诱导细胞分化 ,将细胞周期阻滞于G0 G1 期 ;ATRA下调CDK6、CDK4、cyclinD3和cyclinD1mRNA的水平 ;NaB下调CDK2、cyclinD2和cyclinD1mRNA的水平 ;两药联用下调CDK6、CDK4、CDK2、cyclinD1、cyclinD2和cyclinD3mRNA的水平 ;ATRA和 (或 )NaB均上调P2 1mRNA的水平。结论NaB诱导SKM 1的分化可能是通过上调P2 1mRNA的水平和抑制cyclinD CDK复合体的形成完成的 ,NaB与ATRA对SKM

The study was purposed to explore the molecular mechanisms of sodium butyrate (NaB) action on SKM-1 cell proliferation/differentiation and to study its synergistic effect with all-trans retinoic acid (ATRA). SKM-1 cells were g ro wn in the absence or presence of NaB and/or ATRA; the percentage of viable cells was determined by trypan blue exclusion; differentiation was investigated by ni tro-blue tetrazolium (NBT) reduction; adhesion molecules of cell surface were analysed by FACS; cell cycle distribution was studied after DNA staining by prop idium iodide; D-type cyclins, CDK and P21 mRNA were detected by reverse transcr i ption-polymerase chain reaction (RT-PCR). The results showed that NaB and/or A TRA blocked cells mainly i n the G 0/G 1 phase of the cell cycle; ATRA inhibited the mRNA expression of C DK6, CDK4, cyclin D3 and cyclin D1; NaB inhibited the mRNA expression of CDK2, cycli n D2 and cyclin D1; ATRA and NaB inhibited the mRNA expression of CDK6, CDK4, CD K2, cyclin D1, cyclin D2 and cyclin D3; ATRA and/or NaB both stimulated p21 expr ession at the mRNA levels. It is concluded that the NaB effect on cell proliferation/di fferentiation may be linked to its ability to induce expression of p21 mRNA and inhibit the cyclin D-CDK complexes. These observations support the claim that N aB has the synergistic effect with ATRA.