三氧化二砷和全反式维甲酸联合用药诱导NB4细胞C/EBPε mRNA和CD11b表达的初步研究

The study of C/EBPε mRNA and CD11b expression induced by arsenic trioxide combined with all-trans retinoic acid in NB4 cells

目的 :研究AS2 O3 和AS2 O3 加tRA对NB4细胞凋亡和分化的影响 ,以及NB4细胞CD11b、膜联蛋白 (AnnexinⅤ )和C/EBPεmRNA表达和相互调变关系。方法 :取 1× 10 6个NB4细胞 ,每孔中加入AS2 O3 并同时在另外孔加入AS2 O3 加tRA ,分别在 0、4、8、12、16、2 0、2 4、4 8、72h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AnnexinⅤ ,半定量RT PCR检测C/EBPε表达。结果 :72h内AS2 O3 和AS2 O3 加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。AS2 O3 加tRACD11b的表达增高更加明显,凋亡发生在早期 ,在 4～ 2 0h之内AnnexinⅤ表达较高 ,2 4h后随时间的延长阳性率下降。结论 :AS2 O3 加tRA、AS2 O3 均能诱导CD11b、C/EBPε表达增强 ,可诱导NB4细胞凋亡。在一定剂量下AS2 O3 加tRA具有协同作用 ,诱导NB4细胞分化能力比AS2 O3

Objective:To study the effect of AS 2O 3+tRA and AS 2O 3 on apoptosis and differentiation of NB4 cells so as to evaluate the possible relation of these expressions among CD11b, AnnexinⅤand C/EBPε mRNA.Method:AS 2O 3+tRA and AS 2O 3 were added to the culture system to induce NB4 cells apoptosis and differentiation. cells were incubated and collected at 0, 4, 8, 12, 16, 20, 24, 48 and 72 hours.The expression of CD11b,AnnexinⅤwere determined by FACS.Semi-quantitative RT-PCR was used to detect mRNA expressions of C/EBPε.Result:The mRNA expression of C/EBPε were markedly increased after NB4 stimulated by AS 2O 3+tRA and AS 2O 3 cells within 0 to 72 hours, which leads to NB4 cells CD11b expression increased. The CD11b expression was more significantly increased by AS 2O 3+tRA.Apoptosis was started in the early time after NB4 cells cultured with AS 2O 3+tRA and AS 2O 3.The expression of AnnexinⅤ peak was occurred from 4 to 20 hours. With time prolong the expression of AnnexinⅤ decreased.Conclusion:AS 2O 3+tRA and AS 2O 3 could stimulate NB4 cells expression CD11b, C/EBPε, leaded to NB4 cell apoptosis .In some given dose the effect of AS 2O 3+tRA was more significant to induce NB4 cells apoptosis and differentiation than that of AS 2O 3.