三氧化二砷对尤文肉瘤细胞迁移和侵袭能力抑制作用的体外研究

 目的　研究三氧化二砷在体外对尤文肉瘤细胞系迁移和侵袭能力的抑制作用 。方法　通过噻唑蓝（MTT）实验选择对尤文肉瘤细胞系（A-673、RD-ES）无毒性的三氧化二砷浓度（＜2 μmol／L）后，Transwell迁移和侵袭实验检测三氧化二砷作用下尤文肉瘤细胞的迁移和侵袭能力。Western blot实验及明胶酶谱实验检测PI3K-AKT通路相关作用蛋白及基质金属蛋白酶-9（MMP-9）的表达变化。结果　迁移和侵袭穿过Transwell基底膜的尤文肉瘤细胞的数量随着三氧化二砷浓度的增加而逐渐降低，经不同浓度三氧化二砷处理过的A-673迁移平均数量分别占未处理组的54.3 %、49.0 %和17.0 %，差异有统计学意义（F＝112.78，P＜0.01），在侵袭实验中分别为52.7 %、32.3 %和10.3 %，差异有统计学意义（F＝183.76，P＜0.01）；经过不同浓度三氧化二砷处理后的RD-ES细胞迁移平均数量分别占未处理组的46.0 %、39.0 %和8.0 %，差异有统计学意义（F＝408.25，P＜0.01），在侵袭实验中分别为58.7 %、22.3 %和9.0 %，差异有统计学意义（F＝373.25，P＜0.01）。PI3K-AKT通路相关蛋白和MMP-9表达水平下降。结论　低浓度三氧化二砷可以通过下调PI3K-AKT信号通路的表达来抑制尤文肉瘤细胞系的转移能力。

Inhibition of arsenic trioxide for the capability of migration and invasion in Ewing' s sarcoma cell in vitro

Objective To investigate the effect of arsenic trioxide （As2O3） on the metastasis capability of Ewing＇s sarcoma cells. Methods MTF assays were performed to choose appropriate concentrations of As2O3 （〈 2 μmol/L） for the experiments. Migration and invasion assays were performed to assess the effect of As2O3 on the metastasis of Ewing＇s sarcoma cells. Changes in matrix metalloproteinase （MMP）-9 expressions were detected by gel zymography assay and the phosphoinositide 3-kinase/AKT （PI3K-AKT） pathway was investigated using Western blot. Results The amount of Ewing＇s sarcoma cells across basal membrane of Transwell in migration and invasion assay decreased gradually with the increase in As2O3 concentration. The average quantities of A-673 across the membrane after treatment by gradual concentrations accounted for 54.3 %, 49.0 % and 17.0 % of that of untreated group respectively in migration assay （F =112.78, P 〈 0.01）, while 52.7 %, 32.3 % and 10.3 % in invasion assay （F = 183.76, P 〈 0.01）. Similarly, the percentage of RD-ES was 46.0 %, 39.0 % and 8.0 % in migration assay （F = 408.25, P 〈 0.01） and 58.7 %, 22.3 % and 9.0 % in invasion assay （F = 373.25, P 〈 0.01） respectively. The difference had statistics significance. The expression of MMP-9 was suppressed by As2O3 treatment according to gel zymography assay. Western blot assay s.howed that PI3K-AKT pathway was inhibited and nuclear factor kappa B （NF-kB） was inactivated. Conclusion Low-concentration As2O3 may inhibit metastasis capability of Ewing＇s sarcoma cells.