Department of Oral & Maxillofacial Surgery, Affiliated No. 9 People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
Oral Dis. 2011 May;17(4):362-9. doi: 10.1111/j.1601-0825.2010.01758.x. Epub 2010 Oct 28.
The aim of this study was to examine the anti-invasion effect of Shikonin on human high-metastatic adenoid cystic carcinoma (ACC-M) cells and to explain the possible molecular mechanism involved.
The ACC-M cells were treated with Shikonin (0, 2.5, 5, 10 μM) for 24 h. The protein levels and gelatinolytic activities of MMP-2 and MMP-9 were analyzed using Western blot and Gelatin zymography test, respectively. Matrigel invasion assays were used to investigate tumor invasive potential and electromobility shift assays were used to determine the activity of NF-κB.
The invasiveness of ACC-M cells was reduced in a dose dependent manner following 24-h treatment of up to 10 μM of the Shikonin at which concentration no cytotoxicity occurred. The protein levels and gelatinolytic activities of MMP-9 were significantly suppressed by increasing Shikonin concentrations. The down-regulation of MMP-9 appeared to be via the inactivation of NF-κB as the treatment with Shikonin suppressed the protein level of phosphate-IkBa, which was accompanied by a decrease in DNA-binding level of the factor.
Shikonin inhibits tumor invasion via downregulation of MMP-9 expression in ACC-M cells. Pharmacologic inhibition of the NF-κB-mediated MMP-9 expression by Shikonin might be a powerful treatment option for ACC patients in future.
本研究旨在探讨紫草素对人高转移性腺样囊性癌（ACC-M）细胞的抗侵袭作用，并探讨其可能的分子机制。
用紫草素（0、2.5、5、10 μM）处理 ACC-M 细胞 24 小时。用 Western blot 和明胶酶谱试验分别分析 MMP-2 和 MMP-9 的蛋白水平和明胶酶活性。用 Matrigel 侵袭试验研究肿瘤侵袭潜力，用电迁移率变动分析测定 NF-κB 的活性。
在浓度高达 10 μM 的紫草素作用 24 小时后，ACC-M 细胞的侵袭性呈剂量依赖性降低，而在该浓度下无细胞毒性。随着紫草素浓度的增加，MMP-9 的蛋白水平和明胶酶活性明显受到抑制。MMP-9 的下调似乎是通过 NF-κB 的失活实现的，因为紫草素处理抑制了磷酸化 IkBa 的蛋白水平，同时 NF-κB 的 DNA 结合水平也降低。
紫草素通过下调 ACC-M 细胞中 MMP-9 的表达抑制肿瘤侵袭。紫草素抑制 NF-κB 介导的 MMP-9 表达可能是未来治疗 ACC 患者的一种有效治疗选择。
