Department of Pathology, School of Medicine, Keio University, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-0016, Japan.
Br J Cancer. 2011 Nov 8;105(10):1615-24. doi: 10.1038/bjc.2011.431. Epub 2011 Oct 20.
Although matrix metalloproteinases (MMPs) are implicated in tumourigenesis and cancer progression, the role of MMP-13 in melanoma cell metastases is poorly understood.
Lung metastases of mouse melanoma B16BL6 cells were analysed in MMP-13 knockout (KO) and wild-type (WT) mice after intravenous injection. The mRNA and protein expression of MMP-13 in lung tissues was analysed by RT-PCR, real-time PCR, immunoblotting and immunohistochemistry. The expression of SDF-1α, CXCR4 and endostatin, and effects of endostatin to cultured melanoma cells and lung metastases were also studied.
Lung metastases of B16BL6 cells were significantly higher by 2.5-5.7-fold in MMP-13 KO mice than in WT mice. The expression of MMP-13 in WT mouse lung tissue was stimulated on day 1 after intravenous injection of the melanoma cells and MMP-13 was immunolocalised to vascular endothelial cells in the lungs. Endostatin formation, but not degradation of SDF-1α, in the lung tissue was associated with reduced lung metastasis in WT mice. Endostatin significantly inhibited migration of B16BL6 cells in monolayer wounding assay and remarkably suppressed Matrigel invasion and transendothelial invasion of the cells. In addition, lung metastases of melanoma cells in MMP-13 KO mice were reduced by intraperitoneal administration of endostatin.
Our results suggest that MMP-13 is overproduced by endothelial cells in the lungs with melanoma cells and has a protective role in lung metastasis by local generation of endostatin.
尽管基质金属蛋白酶(MMPs)与肿瘤发生和癌症进展有关,但 MMP-13 在黑色素瘤细胞转移中的作用仍知之甚少。
在静脉注射后,分析 MMP-13 敲除(KO)和野生型(WT)小鼠中的小鼠黑色素瘤 B16BL6 细胞的肺转移。通过 RT-PCR、实时 PCR、免疫印迹和免疫组织化学分析肺组织中 MMP-13 的 mRNA 和蛋白表达。还研究了 SDF-1α、CXCR4 和内皮抑素的表达以及内皮抑素对培养的黑色素瘤细胞和肺转移的影响。
B16BL6 细胞的肺转移在 MMP-13 KO 小鼠中比 WT 小鼠高 2.5-5.7 倍。WT 小鼠肺组织中 MMP-13 的表达在静脉注射黑色素瘤细胞后第 1 天被刺激,并且 MMP-13 被免疫定位到肺部的血管内皮细胞。肺组织中内皮抑素的形成,而不是 SDF-1α 的降解,与 WT 小鼠中肺转移的减少有关。内皮抑素显著抑制 B16BL6 细胞在单层划痕试验中的迁移,并显著抑制细胞在 Matrigel 中的侵袭和穿内皮侵袭。此外,MMP-13 KO 小鼠中黑色素瘤细胞的肺转移通过腹腔内给予内皮抑素而减少。
我们的结果表明,MMP-13 由带有黑色素瘤细胞的肺部内皮细胞过度产生,并通过局部产生内皮抑素来发挥保护作用,从而减少肺转移。