医学论文-三氧化二砷对阿霉素耐药的白血病细胞血管新生相关因子的影响.docVIP

医学论文-三氧化二砷对阿霉素耐药的白血病细胞血管新生相关因子的影响 ???????????????? 作者：陈牧 张建东 张育 王晓铃 沈维干 李国青 【摘要】? 目的探讨三氧化二砷(arsenic trioxide, As2O3)对白血病耐药细胞株K562/A02细胞血管新生相关因子血管内皮生长因子（VEGF）表达水平及基质金属蛋白酶2、9（MMP2、9）活性的影响。 方法用噻唑蓝法(MTT)测定As2O3对K562/A02细胞的增殖抑制作用，用酶联免疫吸附试验测定细胞上清中VEGF含量，用明胶酶谱法半定量测定细胞上清中MMP2、9的活性。 结果K562/A02细胞的IC50是（1.64±0.16）μM，0.05μM As2O3对细胞增殖和VEGF的表达没有明显影响，对MMP2、9的活性在24、48h无明显影响(P0.05)，但在72h则对MMP2、9的活性有明显抑制作用(P0.05)；0.4和3.2μM As2O3可显著抑制细胞增殖，下调VEGF并使MMP2、9活性减弱(P0.05)。结论As2O3可能通过下调VEGF的表达和抑制MMP2、9活性而发??抗白血病细胞血管新生作用。 【关键词】? 三氧化二砷； 血管内皮生长因子； 基质金属蛋白酶； 明胶酶谱 　　Effect of Arsenic Trioxide on Related? Factors of Angiogenesis in K562/A02 Cells 　　Abstract：Objective To investigate the effect of arsenic trioxide (As2O3) on the expression of VEGF and the activity of MMP2 and 9 in K562/A02 cells. MethodsMTT assay was used to detect the inhibition ratio of K562/A02 cell. Enzymelinked immunosorbent assay (ELISA) was used to detect the level of VEGF. Gelatin zymography assay was used to assess the activity of MMP2, 9. ResultsNeither the proliferation nor VEGF expression of K562/A02 cells was influenced by 0.05μM? As2O3 after 24, 48 and 72 hours’ incubation. The activities?? of?? MMP2, 9 were not effected after 24, 48 hours’ incubation with 0.05μM As2O3 (P0.05),? but they were significantly inhibited after 72 hours (P0.05). 0.4 and 3.2μM? As2O3 highly inhibited the proliferation, down regulated VEGF expression and inhibited the activity of MMP2, 9 of K562/A02 cells (P0.05). ConclusionThe possible mechanism of antileukemic angiogenesis by As2O3 may be achieved by inhibiting the expression of VEGF and the activity of MMP2 and 9. 　　Key words：Arsenic trioxide;VEGF; MMP;Gelatin zymography assay 　　0引言 　　肿瘤细胞的多药耐药(Multidrug resistance,? MDR)是导致临床白血病患者化疗失败的常见原因。 造成肿瘤细胞MDR的因素众多， 其中血管新生(Angiogenesis)与肿瘤多药耐药的关系正逐渐受到越来越多的重视。 肿瘤血管新生不仅是白血病发生、 发展和浸润的重要条件， 而且其相关因子， 如血管内皮生长因子(Vascular endothelial growth factor， VEGF)和基质金属蛋白酶(Matrix? metalloproteinase， MMP)所具有的复杂的生物学效应也直接或间接增强了肿瘤细胞的MDR[1, 2]。 因此， 如果能有效下调多药耐药细