2，3-吲哚醌对神经母细胞瘤细胞 SH-SY5Y侵袭转移的影响及机制

摘要

Objective To investigate the effect of 2,3-isatin on the invasion and metastasis of neuroblastoma SH-SY5Y cells and the mechanism.Methods SH-SY5Y cells cultured in vitro were treated for 48 h with different concentra-tions of 2,3-isatin (25,50,100,200 and 300 μmol/L).The cell viability was measured by CCK-8.The number of cells with autolysosome in the control group and 200 μmol/L isatin-treated group was detected by acridine orange staining fluo-rescence.The expression of LC3 Ⅱ,a marker protein for autophagy was examined by Western blotting.The invasion of SH-SY5Y cells was tested by Transwell assay and the motile ability was detected by Scratch test.Results Compared with the control group,the A450 value decreased and the inhibition rate increased when the concentration of 2,3-isatin was higher than 100 μmol/L (P <0.05 or P <0.01).AO staining showed that the number of autolysosomes increased in the 200 μmol/L 2,3-isatin treatment group as compared with that of the control group (P <0.01).Western blotting showed that the expression of LC3-Ⅱincreased with the increasing drug concentration (P <0.05).Transwell assay and Scratch test showed that the invasion and motile abilities of SH-SY5Y cells diminished with the increasing concentration of 2,3-isatin (all P <0.05).Conclusion 2,3-isatin may inhibit the invasion and metastasis of SH-SY5Y cells by inducing the auto-phagy.%目的：探讨2，3-吲哚醌对神经母细胞瘤细胞 SH-SY5Y 侵袭转移的影响及机制。方法体外培养 SH-SY5Y 细胞，用不同浓度（50、100、200、300μmol／L）的2，3-吲哚醌处理细胞48 h，并设不加药物处理的细胞作为对照组，CCK-8法检测细胞活性，筛选合适的2，3-吲哚醌浓度；用吖啶橙染色法检测对照组和200μmol／L 2，3-吲哚醌处理组（加药组）的自噬溶酶体细胞数量；将细胞分别用50、100、200、300μmol／L 2，3-吲哚醌处理（加药组），并设空白对照，48 h 后用 Western blotting 法检测自噬标志性蛋白 LC3-Ⅱ的表达水平，Transwell 小室试验检测细胞的侵袭能力，划痕试验检测细胞迁移能力。结果与对照组相比，随着2，3-吲哚醌浓度的升高，细胞 A450值降低，药物对细胞的抑制率升高（P ＜0．05或＜0．01）。吖啶橙染色结果显示，与对照组相比，加药组产生自噬溶酶体的细胞数增多（P ＜0．01）。Western blotting 结果显示，与空白对照比较，随着药物浓度增加，加药组 LC3-Ⅱ表达增多（P＜0．05或＜0．01）；Transwell 小室试验和划痕试验表明，随着2，3-吲哚醌浓度的增加，SH-SY5Y 细胞侵袭转移能力逐渐减弱（P 均＜0．05）。结论2，3-吲哚醌可能通过诱导神经母细胞瘤细胞 SH-SY5Y 自噬，从而抑制其侵袭转移。