丹参酮Ⅱ_A与ATRA诱导NB4细胞分化过程中黏附分子的变化研究

Variance of Adhesion Molecules in NB4 Cells Differentiation Induced by Tanshinone Ⅱ_A and All-trans Retinoic Acid

目的:研究丹参酮ⅡA(TanⅡA)磺酸钠与全反式维甲酸(ATRA)诱导人急性早幼粒细胞白血病细胞(NB4细胞)分化过程中黏附分子CD44、CD54的变化。方法:取对数生长期的NB4细胞,分为6组,即阴性对照组(培养液中含0.1g.L-1DMSO)和单独作用组1、2(培养液中分别含10μg.L-1TanⅡA磺酸钠、0.5μg.mL-1ATRA)及联合作用组1、2、3(10μg.mL-1TanⅡA磺酸钠分别联合0.5、0.25、0.125μg.mL-1ATRA),均培养5d,计算细胞生长抑制率,观察细胞形态学变化,采用硝基蓝四氮唑还原实验计算阳性细胞率,运用流式细胞术检测黏附分子CD44、CD54的表达。结果:TanⅡA磺酸钠、ATRA能显著提高NB4细胞生长抑制率、诱导分化、增强NBT还原能力(P<0.05);TanⅡA磺酸钠处理后的黏附分子CD44、CD54的表达无显著性改变(P<0.05),ATRA处理后的黏附分子CD44表达减弱、CD54表达增强(P<0.05);24、72、120h之间比较,黏附分子CD44、CD54的表达有显著性差异(P<0.05),但对ATRA(0.125～0.5μg.mL-1)的剂量依赖性不显著(P>0.05)。结论:TanⅡA磺酸钠对NB4细胞的黏附分子CD44、CD54无显著性影响,而ATRA能减弱CD44表达、增强CD54表达。

OBJECTIVE： To investigate the variance of adhesion molecules CD44 and intercellular adhesion molecule 1 （CD54） in human acute promyelocytic leukemia cells （NB4） differentiation induced by tanshinone ⅡA（Tan ⅡA） sulfonate and/ or all- trans retinoic acid （ATRA） . METHODS： The NB4 cells at logarithmic growth phase were assigned to 6 groups： negative control group （containing 0.1 g·L^-1 DMSO in culture fluid）, single action group Ⅰ （containing 10 μg· L^-1 Tan ⅡA sulfonate） ,single action group Ⅱ （containing 0.5 μg ·mL^-1 ATRA）, joint action group 1, 2, and 3 （in which 10μg·L^-1 Tan ⅡA sulfonate was used concomitantly with 0.5, 0.25 and 0.125 μg·mL^-1 ATRA, respectively）, with cells in all groups cultured for 5 days. The growth inhibiting ratio of NB4 cells was computed, the cell morphology change was observed; the positive cell ratio was computed using nitro - blue tetrazoline reduction experiment and the expressions of CD44 and CD54 were detected by flow cytometry .RESULTS： Tan ⅡA sulfonate and ATRA significantly enhanced the growth inhibition ratio of NB4 cells, induced differentiation ability and reduction ability of NBT as compared with the control （P（0.05） . No significant differences were noted for the expression of Tan HA sulfonate treated CD44 and CD54（P〈0.05）, but ATRA significantly inhibited CD44 expression and enhanced CD54 expression of NB4 cells（P 〈 0.05） .The expression of CD44 and CD54 at 24, 72 and 120 h were significantly different （P〈0.05）, but the dose-dependence manner of ATRA （0.125-0.5 tLg· mL^-1） was not significant（P 〉 0.05）. CONCLUSION： The influence of Tan HA sulfonate on the expression of CD44 and CD54 of NB4 cells was nonsignificant, but ATRA could attenuate CD44 expression and enhance CD54 expression of NB4 cells.