As2O3 不同给药方式对 NB4 细胞侵袭力和 MMPs 活性的影响

 目的 对比 As₂O₃ 不同干预方式对白血病细胞株 NB4 的侵袭力和基质金属蛋白酶 2,9(MMP-2 、 MMP-9) 活性的影响。 方法 噻唑蓝（ MTT ）法检测 As₂O₃ 恒定和变化两种体系体外干预对 NB4 细胞的增殖抑制， ELISA 检测 2 种体系中处理的 NB4 细胞的 MMP-2 和 MMP-9 蛋白含量的变化。明胶酶谱法检测 2 种 As₂O₃ 处理前后 NB4 细胞 MMP-2 活性变化， Westen blot 检测活性 MMP-9 蛋白表达， Transwell 小室穿透实验检测两种 As₂O₃ 处理方式对 NB4 细胞穿透力的影响。 结果 ①与平行对照组比较 0.1 μmol·L^-1 As₂O₃ 恒定浓度体系中持续作用 72 h ，对 MMP-2 、 MMP-9 的活性和 NB4 细胞侵袭力影响不显著。 0.5 μmol·L^-1 As₂O₃ 持续作用 24 h 与对照组比较无明显差异， 48 h 以后 MMP-2 、 MMP-9 的活性和 NB4 侵袭力降低，与对照组比较差异显著。 1.0 μmol·L^-1 以上浓度 As₂O₃ 处理 24 h ， MMP-2 、 MMP-9 活性和 NB4 细胞侵袭力下降，并呈时间依赖性。② 2.0 μmol·L^-1 As₂O₃ 恒定浓度体系与峰浓度为 5.0 μmol·L^-1 ，终质量浓度为 0 μmol·L^-1 的变化浓度体系比较，前者对 NB4 的侵袭力和 MMPs 活性的抑制程度比后者更显著。 结论 NB4 细胞的侵袭力与其 MMP-2 、 MMP-9 活性有关，促分化有效的低浓度 As₂O₃ 对 NB4 细胞侵袭力和 MMP-2 、 MMP-9 活性抑制作用不明显。在干预时间相同的情况下，促凋亡有效的恒定浓度 As₂O₃ 持续作用，对 NB4 细胞侵袭力和 MMP-2 、 MMP-9 活性的抑制强度大于峰浓度很高但持续时间很短的变化浓度体系 , 这可能是临床上 As₂O₃ 持续缓慢输注法较常规给药法更能有效降低急性早幼粒细胞白血病（ APL ）中枢神经系统浸润和早期的致死性脑出血发生率的机制之一。

Influence of Two Arsenic Trioxide Treatment Regimen on Activity of MMPs and Cell Invasion in NB4 Cells

OBJECTIVE To compare the influence of two kinds arsenic trioxide (As2O3) treatment on activity of MMPs and cell invasion in NB4 cells. METHODS NB4 cell line were treated with steady and changing concentrations of As2O3 in vitro respectively. The cell death was investigated by MTT assay. The activities of MMPs were detected by enzyme-linked immunosorbent assay, and their expressions were measured by western blotting and gelatinase zymography. The migration capabilities of NB4 at baseline and after being treated with the two kinds of different As2O3 treatment were evaluated by transwell assay. RESULTS ①With As2O3 continuous treatment in steady concentration: the MMPs activity and migration of NB4 cells were not significantly decreased by 0.1 μmol·L-1 As2O3 for 72 h. The activity was remarkably inhibited by As2O3 in 0.5 μmol·L-1 for 72 h, in 1.0 μmol·L-1 for 48 h and in 2.0 μmol·L-1 and above 24 h compared with the parallel control. ②The inhibition effects of As2O3 on MMPs activity and migration of NB4 with As2O3 in 2.0 μmol·L-1 steady concentration treatment were remarkable compared with that in changing concentration treatment in vitro, even though the peak of As2O3 concentration was 5.0 μmol·L-1. CONCLUSION As2O3 with lower concentration could not inhibit MMPs activity and invasion of NB4 effectively. however, As2O3 at higher apoptosis effective concentration decreased the MMPs activity and inhibited NB4 invasion remarkably in a concentration- and time-dependent manner. The inhibition efficiency of As2O3 on MMPs activity and NB4 cell invasion treated with general speed As2O3, which has a fluctuated serum arsenic and a long pause of apoptosis non-effective lower arsenical level in serum between the two dosages of As2O3 administration, was less than that treated with the slow-steady speed As2O3 infusion, which can keep a relatively constant promoting apoptosis effective level of serum arsenic during the whole As2O3 infusion treatment.