人单核细胞白血病细胞系SHI-1的诱导分化和凋亡

  目的 对本实验室建立的一株新的人单核细胞白血病细胞系SHI-1进行诱导分化和凋亡的研究。方法 以瑞特染色、四唑氮蓝还原试验（NBT）、流式细胞仪（FCM）检测Annexin V以及CD11b和CD14表达的改变后和DNA凝胶电泳等方法观察三丁酸甘油酯（TB）作用后SHI-1细胞的改变；以瑞特染色、FCM检测Annexin V以及CD11b和CD14表达的改变等方法观察三氧化二砷（As2O3）作用后SHI-1细胞的改变。结果 SHI-1细胞经TB处理后，可出现典型的凋亡形态学改变，部分细胞出现部分分化，NBT实验显示还原率上升，FCM结果示CD11b的表达无改变，而CD14的表达上调，Annexin V的检测结果显示SHI-1细胞经TB作用24 h后凋亡率明显上升，可见典型的DNA梯带。SHI-1细胞经As2O3作用后可见典型的凋亡形态学改变，部分细胞可见部分分化的改变，SHI-1细胞CD14的表达上调，CD11b的表达无改变，FCM检测Annexin V显示凋亡率明显上升。TPA作用24 h后SHI-1细胞基本贴壁，形态不规则，有伪足形成。结论 TB对SHI-1细胞具有诱导分化和凋亡的双向调节作用。TPA可诱导SHI-1细胞分化和贴壁。0.5 μmol／L、1.0 μmol／L和2.0 μmol／L As2O3可诱导SHI-1细胞发生凋亡和部分分化。

Preliminary study of inducing differentiation and apoptosis in a novel human acute monocytic leukemic cell line SHI-1

Objective To explore the inducing differentiation and apoptosis in a novel human acute monocytic leukemic cell line SHI-1. Methods Cell morphological analysis, NBT reduction test, the expres－sion of CD11b and CD14, cell cycle analysis and binding of annexinV tested by FCM, DNA electrophoresis were performed to evaluate whether tributyrin (TB), arsenic trioxide (As2O3), and 12-O-tetradecanoyl-phorbol-13 acetate (TPA) could induce the differentiation and apoptosis in the SHI-1 cell line. Results The results of morphological analysis, NBT reduction test, and FCM assay showed that TB could induce differentiation of SHI-1 cells. The SHI-1 cell line presented typical apoptotic evidences including morphology features, DNA ladders and the increased apoptotic rate determined by FCM. Moreover, the SHI-1 cell line presented typical apoptotic morphology after being treated with As2O3. The apoptotic rate of the SHI-1 cell line treated with As2O3 was increased determined by FCM. The expression level of CD14 was upregulated while that of CD11b was unchanged in the SHI-1 cell line. Most of the SHI-1 cells adhered to the flask after being treated with TPA for 24 hours. Conclusions TB may produce dual effects of differentiation and apoptosis. The cell line could be induced to differentiation partially or apoptosis after being treated with 0.5 μM, 1.0 μM, and 2.0 μM As2O3. TPA could induce differentiation and adhesion of the SHI-1 cell line.