CD44V6纳米抗体诱导NB4细胞分化的作用与机制研究

The effect and mechanism of CD44V6 nanobody on inducing differentiation of NB4 cells

目的:探讨CD44V6纳米抗体诱导NB4细胞分化的作用与分子机制。方法:采用CCK8细胞增殖试剂盒观察CD44V6纳米抗体对NB4细胞增殖的影响,流式细胞仪检测细胞周期的变化,流式细胞术检测分化抗原CD11b的表达变化,Western blotting检测细胞c-Myc的蛋白表达变化。结果:1 ng/mL的CD44V6纳米抗体于第3天发生NB4细胞的增殖抑制作用,而在10 ng/mL时,第1天时即能发生细胞增殖抑制作用。在10 ng/ml的浓度诱导条件下,体外共培养诱导3天后,NB4细胞周期变化的特点是G0/G1期明显增高,S期明显降低,G2/M期也呈现降低趋势。NB4细胞的分化抗原CD11b表达升高,c-Myc蛋白的表达明显降低。结论:CD44V6纳米抗体诱导NB4细胞分化成熟的作用可能与c-Myc的表达水平降低有关。

Objective:To investigate the effect and molecular mechanism of CD44V6 nanobody on the differentiation of NB4 cells.Methods:CCK8 cell proliferation kit was used to observe the proliferation of NB4 cells,flow cytometry was used to detect cell cycle profile,flow cytometry was used to detect the expression of CD11b antigen,and Western blotting was used to detect c-Myc expression.Results:The 1ng/mL CD44V6 Nanobody inhibited the proliferation of NB4 cells on the 3rd day,and 10 ng/mL CD44V6 Nanobody could inhibit the cell proliferation of NB4 cells even on the 1st day.After 3 days of in vitro co-culture by exposure to 10 ng/ml CD44V6 Nanobody,NB4 cell cycle profile was characterized by a significant increase in the G0/G1 phase,a significant decrease in the S phase,and a decreasing trend in the G2/M phase.The expression of the differentiation antigen CD11b of NB4 cells was increased,and the expression of c-Myc protein was significantly decreased.ConClusion:The effect of CD44V6 Nanobody on inhibiting the proliferation of NB4 cells and inducing their differentiation may be related to the decrease of c-Myc expression.