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基质金属蛋白酶在大黄素抑制人神经母细胞瘤SH-SY5Y细胞侵袭和迁移中的作用

Involvement of matrix metalloproteinases on the inhibition of cells invasion and migration by emodin in human neuroblastoma SH-SY5Y cells.

作者信息

Lu Hsu-Feng, Lai Kuang-Chi, Hsu Shu-Chun, Lin Hui-Ju, Kuo Chao-Lin, Liao Ching-Lung, Yang Jai-Sing, Chung Jing-Gung

机构信息

Department of Clinical Pathology, Cheng Hsin Rehabilitation Medical Center, Taipei, Taiwan, ROC.

出版信息

Neurochem Res. 2009 Sep;34(9):1575-83. doi: 10.1007/s11064-009-9946-3. Epub 2009 Mar 17.

Abstract

Emodin (1,3,8-trihydroxy-6-methylanthaquinone), an active component present in the root and rhizome of Rheum palmatum L. (Polygonaceae) has anti-bacterial, anti-tumor, diuretic and vasorelaxant effects. However, its mechanism of action on the cell migration and invasion of human neuroblastoma cancer SH-SY5Y cells is not fully understood. In this study, firstly, the effects of emodin on the percentage of viable cells were examined by using MTT assay and it was found that emodin induced dose-and time-dependent inhibition in human neuroblastoma SH-SY5Y cells. Second, the effects of emodin on the migration and invasion of SH-SY5Y cells were examined by using wound assay and matrigel counting and the results showed that emodin suppressed the migration and invasion of SH-SY5Y cells. Third, we examined the effect of emodin on the levels of associated proteins by using Western blotting and the results indicated that emodin inhibited the levels of GRB2, RhoA, HIF-1alpha, VEGF, FAK, iNOS, COX2, p-p38, p-c-jun, MMP2, MMP9 and MMP7 but promoted the levels of PKC, PI3K, MEKK3 and NF-kappaB p65 that led to the inhibition of migration and invasion of SH-SY5Y cells in vitro.

摘要

大黄素(1,3,8 - 三羟基 - 6 - 甲基蒽醌)是掌叶大黄(蓼科)根和根茎中的一种活性成分,具有抗菌、抗肿瘤、利尿和血管舒张作用。然而,其对人神经母细胞瘤SH - SY5Y细胞迁移和侵袭的作用机制尚未完全明确。在本研究中,首先,通过MTT法检测大黄素对活细胞百分比的影响,发现大黄素对人神经母细胞瘤SH - SY5Y细胞具有剂量和时间依赖性抑制作用。其次,采用划痕实验和基质胶计数法检测大黄素对SH - SY5Y细胞迁移和侵袭的影响,结果表明大黄素抑制SH - SY5Y细胞的迁移和侵袭。第三,通过蛋白质免疫印迹法检测大黄素对相关蛋白水平的影响,结果显示大黄素抑制GRB2、RhoA、HIF - 1α、VEGF、FAK、iNOS、COX2、p - p38、p - c - jun、MMP2、MMP9和MMP7的水平,但促进PKC、PI3K、MEKK3和NF - κB p65的水平,从而导致体外SH - SY5Y细胞迁移和侵袭受到抑制。

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