三氧化二砷和全反式维甲酸联合使用诱导NB4细胞C/EBPεmRNA的表达

C/ EBP ε mrNA Expression Profile Induced by Arsenic Trioxide Combined with all-trans Retinoic acid in NB4 Cells

背景与目的:研究三氧化二砷+全反式维甲酸(As2O3+tRA)和As2O3对NB4细胞凋亡和分化的影响,NB4细胞CD11b、Annexin Ⅴ和C/EBP ε、PML-RAR α基因mRNA表达和相互调变关系。材料与方法:取106个NB4细胞,每孔中加入As2O3并同时在另外孔加入As2O3+tRA,分别在0、4、8、12、16、20、24、48、72 h时取出细胞,用流式细胞术检测细胞分化指标CD11b和凋亡指标Annexin Ⅴ,半定量RT-PCR检测C/EBPε和PML-RAR α基因的表达。结果:72 h内As2O3和As2O3+tRA均能诱导NB4细胞C/EBPε表达增强4倍,从而进一步诱导CD11b的表达。As2O3+tRA CD11b的表达增高更加明显,As2O3和As2O3+tRA两组药物均有显著的降解PML-RARα基因的作用,As2O3+tRA组PML-RAR α融合基因降解更为明显,凋亡发生在早期,在4-20 h之内Annexin Ⅴ表达较高,24 h后随时间的延长阳性率下降。结论:As2O3+tRA、As2O3均能诱导CD11b、C/EBPε表达增强,可使PML-RAR α融合基因降解,诱导NB4细胞凋亡。在一定剂量下As2O3+tRA具有协同作用,诱导NB4细胞分化和凋亡的能力比As2O3单独用药组更强。

BACKGROUND & AIM: To study the effect of As2O3 + tRA and As2O3 on apoptosis and differentiation of NB4 cells to evaluate the possible relation of these expressions among CD11b, Annexin Ⅴ and C/EBPε, PML-RARαmRNA. MATERIAL AND METHODS: As2O3 + tRA and As2O3 were added to the culture system to induce NB4 cells apoptosis and differentiation. NB4 cells were incubated and collected at 0, 4, 8, 12, 16, 20, 24, 48 and 72 hours , The expression of CD11b, Annexin Ⅴ were determined by FACS, We used semi-quantitative RT -PCR.to detect mRNA expressions of C/EBP ε, PML/RAR α. RESULTS: The mRNA expression of C/EBP ε were markedly increased after NB4 cells stimulated by As2O3 + tRA and As2O3 within 0 to 72 hours, it leads to NB4 cells CD11b expression increased. The CD11b expression was more significant increased by As2O3 + tRA . As2O3 + tRA and As2O3 had the same role to degrade PML-RAR a fuse gene, As2O3 + tRA was more powerful in degra-dating PML-RAR α fuse gene. Apoptosis was started in the early time after NB4 cells cultured with As2O3 + tRA and As2O3 ,The expression of Annexin Ⅴ peak was occurred from 4 to 20 hours, With time prolong the expression of Annexin Ⅴ decreased. CONCLUSION: As2O3 + tRA and As2O3 could stimulate NB4 cells expression CD11b, C/ EBP ε and decrease PML-RAR α fuse gene, leaded to NB4 cells apoptosis .In some given dose the effect of As2O3+tRA was more significant to induce NB4 cells apoptosis and differentiation than that of As2O3.