三氧化二砷抑制神经母细胞瘤细胞侵袭性实验研究

Experimental Study on Effect of Arsenic Trioxide on Suppression of Neuroblastoma Cell Invasion

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 whether arsenic trioxide （ As2O3 ） with different density is capable of affecting the invasiveness of neuroblastoma （NB） cells, and to give grounds for NB therapy with As2O3. Methods 1. Well - developed NB cells were selected and exposed to 0. 75 μmol/L,1.50 μmol/L,3.0 μmol/L As2O3 for 24 h;2. Collect the adherence cells,count the number and float them in nutrient medium again,add them into the transwell polycarbonate membrane plate that was covered by matrigel, there were 2 × 10^4 NB cells in each well; 3. After 24 h, take off the membrane, fix the cells which cross the membrane with mehanol and dye them with hematoxylin;4. Observe the NB cells and count them, so the capability of invasion of LA - N - 5 was evaluated by transwell chamber assay. Results After 24 h with the different density As2O3, the number of invading LA - N -5 cells was significantly lower in As2O3 group than that in control group（ the number of invading cells of the As2O3 group was 28.0 ± 4.0,19.33 ± 4.16,6.33 ± 1.53, respectively, the cell number of the control group was 46.33 ±6.11 ）（P =0. 013,0.003,0） ;among the experiment groups, there was no difference between 0.75 μmol/L and 1.50 μmol/L （ P = 0.06）, and it was significantly different between 0.75 μmol/L and 3.0 μmol/L, 1.50 μmol/L and 3.0 μmol/L （ P = 0,0.007 ）, the number of invading LA - N - 5 cells of 3.0μmol/L As2O3 was the least. Conclusions As2O3 could inhibit the invasive potential of NB cells; the inhibitory action of 3.0 μmol/L As2O3 is the most.