人干细胞标志musashi2在急性髓系白血病细胞分化中的表达.docVIP

人干细胞标志Musashi2在急性髓系白血病细胞分化中的表达 张慧娟，谭 诗，陈莎娜，王 娟，覃凤娴，陈先春，张 伶 （400016重庆重庆医科大学检验临床检验诊断学教育部重点实验室目的 探讨人干细胞标志Musashi2（Msi2）在佛波酯（phorbol myristoyl acetate，PMA）诱导急性单核白血病细胞株THP-1分化过程中的表达。方法 50 n PMA作用THP-1细胞6、12、24 h后，倒置显微镜下观察THP-1细胞贴壁和形态改变；流式细胞仪检测THP-1细胞表面分化抗原CD11b的表达量PCR和 estern blot检测Msi2的蛋白表达水平；定量PCR检测Msi2下游信号 Numb的mRNA水平量PCR和 estern blot检测结果 镜下观察，PMA诱导细胞分化过程中细胞贴壁逐渐增多并开始有明显的形态学改变。流式结果表明CD11b表达随THP-1细胞分化增多而增加。定量PCR和estern blot结果表明，与0 h组比较，PMA处理Msi2的mRNA和蛋表达下（P0.05），Numb表达逐渐增高（P0.05），Notch1表达随THP-1细胞分化显著下降（0.05）。结论 干细胞标志Msi表达随THP-1细胞分化而下降，可能参与白血病细胞分化的过程。 Musashi2；分化； 佛波酯；myelogenous leukemia cell Zhang Huijuan, Tan Shi, Chen Shana, Wang Juan, Qin Fengxian, Chen Xianchun, Zhang Ling (Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China) [Abstract] Objective To determine the expression of Musashi2(Msi2) , a human stem cell marker in the differentiation of human myelomonocytic leukemia cell line THP-1 induced by phorbol myristoyl acetate (PMA). Methods The differentiation of THP-1 cells was induced by 50 nmol/L PMA. Within 0, 6, 12 and 24 hours treatment, The cell adhesion and morphology state were detected by inverted microscope. The expression of CD11b, THP-1 cell differential surface marker was assayed using flow cytometry (FCM). Real-time PCR and Western blotting were employed to test the mRNA and protein expression of Msi2. The expression of Msi2 downstream signal molecule Numb was detected by Real-time PCR. The mRNA and protein expression of Notch1, Numb downstream signal molecule were detected by Real-time PCR and Western blotting too. Results During the differentiation process more cells became adherent and changed morphology significantly. FCM indicated that the expression of CD11b was increased with THP-1 cell differentiation. Real-time PCR and western blotting showed that compared with the 0 h group both of mRNA and protein expression of Msi2 we