甲异靛对急性早幼粒细胞白血病细胞系NB_4体外作用的研究

Studying of effects of meisoindigo on acute promyelocytic leukemia cell line NB_4 cells in vitro

目的探讨甲异靛对NB4细胞系的抑制增生、诱导凋亡和分化的作用。方法用锥虫蓝染色方法进行活细胞计数;用NBT还原实验和CD11a、CD11b的表达判定促分化作用;光镜和电镜细胞形态学、TUNEL标记和Sub-G1测定观察细胞凋亡,RT-PCR和流式细胞术分析法检测凋亡调控基因mRNA及蛋白水平表达。结果20μmol/L和30μmol/L甲异靛可明显抑制NB4细胞的增生,诱导细胞凋亡并下调bcl-2表达,使细胞停滞在G0/G1期。5μmol/L和10μmol/L时NB4细胞的NBT还原能力明显升高,CD11a表达明显升高。结论甲异靛通过诱导细胞凋亡和使细胞停滞在G0/G1期这两种机制显著抑制NB4细胞增生,此外,还有部分诱导细胞分化的作用。

Objective To investigate the effects of meisoindigo on proliferation inhibit, induce apoptosis and differentiation of NB4 cells in vitro. Methods The cell growth rate was tested by trypan blue. The effects of induce differentiation were determined by detection of the NBT reduction value and the expression of CD11a. The effects of induce apoptosis were. determined by morphology, TUNEL labeling and detection of Sub- G1 cell population. The mRNA and protein expression levels of apoptosis regulation genes were detected by RT- PCR and FACS assay. Results 20 μmol/L and 30 μmol/L meisoindigo significantly inhibited the growth of NB4 cells, induced the NB4 cells to apoptosis, decreased the expression of bcl- 2 on mRNA and protein level, and arrested the cells in the G0/G1 phase in concentration- and time- dependent manner. 5 μmol/L and 10 μmol/L meisoindigo significantly increased the NBT reduction value and the expression of CD11a as compared with the control. Conclusions Meisoindigo significantly inhibited the proliferation of NB4 cells through inducing cell to apoptosis and arresting the cells in the G0/G1 phase of the cell cycle, and partially induced differentiation of NB4 cells. [