Osaka Eiji, Yang Xiaoqian, Shen Jacson K, Yang Pei, Feng Yong, Mankin Henry J, Hornicek Francis J, Duan Zhenfeng
Sarcoma Biology Laboratory, Department of Orthopedic Surgery, Massachusetts General Hospital, 50 Fruit Street, Jackson 1115, Boston, Massachusetts, 02114; Department of Orthopaedic Surgery, Nihon University School of Medicine, 30-1 Oyaguchikami-cho, Itabashi-ku, Tokyo, 173-8610, Japan.
J Orthop Res. 2014 Aug;32(8):1075-82. doi: 10.1002/jor.22632. Epub 2014 Apr 24.
Recent studies have revealed that expression of miRNA-1 (miR-1) is frequently down-regulated in several cancer types including chordoma. Identifying and validating novel targets of miR-1 is useful for understanding the roles of miR-1 in chordoma. We aimed to further investigate the functions of miR-1 in chordoma. Specifically, we assessed whether restoration of miR-1 affects cell migration and invasion in chordoma, and focused on the miR-1 potential target Slug gene. Migratory and invasive activities were assessed by wound healing and Matrigel invasion assays, respectively. Cell proliferation was determined by MTT assay. Slug expression was evaluated by Western blot, immunofluorescence, and immunohistochemistry. Restoration of miR-1 expression suppressed the migratory and invasive activities of chordoma cells. Transfection of miR-1 inhibited cell proliferation both time- and dose-dependently in chordoma. MiR-1 transfected cells showed inhibited Slug expression. Slug was over-expressed in chordoma cell lines and advanced chordoma tissues. In conclusion, we have shown that miR-1 directly targets the Slug gene in chordoma. Restoration of miR-1 suppressed not only proliferation, but also migratory and invasive activities, and reduced the Slug expression in chordoma cells. These results collectively indicate that miR-1/Slug pathway is a potential therapeutic target because of its crucial roles in chordoma cell growth and migration.
最近的研究表明，包括脊索瘤在内的几种癌症类型中，miRNA-1（miR-1）的表达常常下调。鉴定和验证miR-1的新靶点有助于理解miR-1在脊索瘤中的作用。我们旨在进一步研究miR-1在脊索瘤中的功能。具体而言，我们评估了miR-1的恢复是否会影响脊索瘤细胞的迁移和侵袭，并聚焦于miR-1的潜在靶点Slug基因。分别通过伤口愈合试验和基质胶侵袭试验评估迁移和侵袭活性。通过MTT试验测定细胞增殖。通过蛋白质免疫印迹、免疫荧光和免疫组织化学评估Slug的表达。miR-1表达的恢复抑制了脊索瘤细胞的迁移和侵袭活性。miR-1转染在脊索瘤中对细胞增殖具有时间和剂量依赖性的抑制作用。转染miR-1的细胞显示出Slug表达受到抑制。Slug在脊索瘤细胞系和晚期脊索瘤组织中过表达。总之，我们已经表明miR-1在脊索瘤中直接靶向Slug基因。miR-1的恢复不仅抑制了增殖，还抑制了迁移和侵袭活性，并降低了脊索瘤细胞中Slug的表达。这些结果共同表明，miR-1/Slug通路因其在脊索瘤细胞生长和迁移中的关键作用而成为一个潜在的治疗靶点。
