三氧化二砷对K562细胞基质金属蛋白酶-2、9活性的影响

目的探讨三氧化二砷(ATO)对白血病K562细胞株基质金属蛋白酶-2、9(MMP-2、MMP-9)活性的影响。方法用0、0.05、0.40和3.20μmol/L的ATO分别处理K562细胞,培养24、48、72 h后收获细胞,对细胞上清进行明胶酶谱分析。结果①MMP-2、9在K562细胞均有表达,其活性条带在对照组以MMP-2(72 kd)最宽,MMP-9次之,MMP-2(62 kd)最窄。②MMP-2、9经0.05μmol/L ATO处理72 h后差异具有显著性(P<0.05);经0.4、3.2μmol/L的ATO处理后随作用时间延长而抑制现象也逐渐增强,与对照组比较均有显著性差异(P<0.05)。结论ATO可以有效抑制K562细胞MMP-2、MMP-9的活性,但其作用的强度与剂量及作用时间有关。ATO对MMP-2、MMP-9的活性的抑制可能是其抗白血病细胞血管新生的机制之一。

EFFECT OF ARSENIC TRIOXIDE ON MMP-2,MMP-9 EXPRESSED IN K562 CELL

Objective To investigate the effect of various concentration of ATO on the activity of MMP-2,MMP-9 expressed in K562 cells treated with ATO.Methods K562 cells were seeded in 24-well plates(1×10~5·mL~(-1)) and harvested after 24,48 and 72 h incubation with 0,0.05,0.4 and 3.2 μmol/L ATO.The activity of MMP-2,MMP-9 was assessed by gelatin zymography assay.Results ①K562 cells expressed both MMP-2 and MMP-9.Among MMP-2(72KD),MMP-2(62KD) and MMP-9 in the control group,the electrophoresis strip of MMP-2(72KD) was the widest,MMP-9 was the second ,and MMP-2(62KD) was the last.②After seeded 72 h,the activity of MMP-2 was inhibited by 0.05 μmol/L ATO(P<0.05).With the increase of ATO incubation time,the inhibited effect was reinforced by 0.4 and 3.2 μmol/L ATO.Conclusion ATO can inhibit the expression of the activity of MMP-2,MMP-9,but it was influenced by the dose and time.One of the possible mechanisms of anti-leukemic angiogenesis by ATO may be achieved by inhibiting the expression MMP-2,MMP-9.