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目的探讨靛玉红甲肟(Indirubin-3’-monoxime,IRO)对人喉癌Hep-2细胞迁移和侵袭的影响及其机制。方法MTT法检测不同浓度、不同作用时间IRO对Hep-2细胞增殖活性的影响。Boyden chamber实验计算各组穿膜的细胞数以比较靛玉红甲肟对细胞侵袭能力的变化。明胶酶谱检测不同浓度靛玉红甲肟处理喉癌Hep-2细胞后细胞MMP-9和MMP-2活性的变化。结果 MTT结果发现IRO对Hep-2细胞具有明显的增殖抑制作用,且表现为剂量依赖性(P<0.01)。Boydenchamber体外迁移和侵袭实验结果显示:靛玉红甲肟可以显著抑制人喉癌Hep-2细胞的侵袭能力(P<0.05)。明胶酶谱检测发现以靛玉红甲肟处理24h后人喉癌Hep-2细胞MMP-9和MMP-2活性明显降低,不同浓度组间存在显著性差异。结论靛玉红甲肟具有抑制人喉癌Hep-2细胞株增殖和侵袭的作用,其作用机制与显著抑制MMP-9和MMP-2活性有关。
Objective To investigate the effect of indirubin-3’-monoxime (IRO) on the migration and invasion of human laryngeal carcinoma Hep-2 cells and its mechanism. Methods MTT assay was used to detect the proliferation of Hep-2 cells with IRO at different concentrations and different time. Boyden chamber experiment calculated the number of cells penetrating the membrane in order to compare the indomethacin-induced changes in cell invasive ability. Gelatin zymography was used to detect the changes of MMP-9 and MMP-2 activity in Hep-2 cells treated with different concentrations of indirubin. Results MTT results showed that IRO had a significant inhibitory effect on Hep-2 cells in a dose-dependent manner (P <0.01). Boydenchamber migration and invasion in vitro showed that indirubin-methyl oxime could significantly inhibit the invasion of human laryngeal carcinoma Hep-2 cells (P <0.05). Gelatin zymography showed that the activity of MMP-9 and MMP-2 in human laryngeal carcinoma Hep-2 cells was significantly decreased after treated with indirubin-methyl oxime for 24 h. There was a significant difference between the different concentrations. Conclusion Indirubinximethromycin can inhibit the proliferation and invasion of human laryngeal carcinoma Hep-2 cells. Its mechanism of action is related to the significant inhibition of the activity of MMP-9 and MMP-2.