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目的探讨不同水平三氧化二砷(As2O3)对神经母细胞瘤(NB)细胞侵袭力的影响,为As2O3用于NB的治疗提供理论依据。方法1.选取生长良好的NBLA-N-5细胞,分别按As2O3终浓度为0.75、1.50、3.0、6.0μmol/L加药,作用24h;2.收集贴壁细胞,计数并重新悬浮,加入到铺有人工基底膜胶的细胞侵袭小室,使每孔约为2×104个细胞,孵育24h;3.取下人工基底膜胶,甲醇固定穿过膜的肿瘤细胞,苏木晶染色;4.光学显微镜观察肿瘤细胞并计数穿过膜的肿瘤细胞数量,了解细胞侵袭力的改变。结果砷剂作用24h后,实验组的LA-N-5侵袭细胞数较对照组显著减少(实验组穿过膜的细胞数分别为28.0±4.0,19.33±4.16,6.33±1.53,对照组平均为46.33±6.11)(P=0.013,0.003,0);实验组0.75μmol/L与1.50μmol/L组间结果比较无显著性差异(P=0.06);0.75μmol/L与3.0μmol/L、1.50μmol/L与3.0μmol/L组间比较均有显著性差异(P=0,0.007),以3.0μmol/L组穿过膜的细胞数最少。结论As2O3能够抑制NB细胞系LA-N-5细胞的侵袭力;3.0μmol/L组的As2O3抑制作用最强。
Objective To investigate the effects of different levels of arsenic trioxide (As2O3) on the invasiveness of neuroblastoma (NB) cells and to provide a theoretical basis for the treatment of NB with As2O3. Methods 1. Select well-growing NBLA-N-5 cells, respectively, according to As2O3 final concentration of 0.75,1.50,3.0,6.0μmol / L dosing for 24h; 2. Adherent cells were collected, counted and resuspended, added to The cells covered with artificial basement membrane glue invade the cells and make about 2 × 104 cells per well and incubate for 24 hours. 3. Remove the artificial basement membrane glue and fix the tumor cells through the membrane with methanol, somatoxylin staining.4. Tumor cells were observed and the number of tumor cells that passed through the membrane was counted to see the change in cell invasiveness. Results After treated with arsenic for 24 hours, the number of invasive cells of LA-N-5 in the experimental group was significantly decreased compared with the control group (the number of cells passing through the membrane in the experimental group was 28.0 ± 4.0, 19.33 ± 4.16 and 6.33 ± 1.53 respectively, 46.33 ± 6.11) (P = 0.013,0.003,0). There was no significant difference between the experimental groups of 0.75μmol / L and 1.50μmol / L (P = 0.06); 0.75μmol / L and 3.0μmol / There was a significant difference between the two groups (P = 0,0.007). The number of cells passing through the membrane was the least in 3.0μmol / L group. Conclusion As2O3 can inhibit the invasiveness of NB cell line LA-N-5 cells. As2O3 at 3.0μmol / L has the strongest inhibitory effect.