余铃，张政佩，孙祥然，郑迪，郭卫春

（武汉大学人民医院骨一科，武汉 430060）

miR-429 通过靶向作用Sox2抑制骨肉瘤干细胞特性

余铃，张政佩，孙祥然，郑迪，郭卫春

（武汉大学人民医院骨一科，武汉 430060）

目的探讨miR-429在骨肉瘤干细胞中的作用。方法利用不同方法筛选出骨肉瘤干细胞，检测miR-429的含量。利用分子探针技术将骨肉瘤细胞分为miR-429高表达组和低表达组，检测各组骨肉瘤细胞干细胞样特性。利用转染技术检测miR-429对骨肉瘤干细胞样特性的影响。利用荧光素酶报告基因系统，验证miR-429下游靶基因。结果骨肉瘤干细胞中miR-429的表达量明显减少。miR-429高表达细胞的干细胞特性明显低于miR-429低表达细胞，且转染实验验证了上述结果。荧光素酶报告实验显示，miR-429在转录后水平上负性调节Sox2基因。结论miR-429可以通过下调Sox2负性调节骨肉瘤干细胞特性。

骨肉瘤； 干细胞； miRNA-429； Sox2

Abstract ObjectiveTo investigate the role of miR-429 in osteosarcoma stem cells.MethodsWe employed different approaches to test the expression of miR-429 in osteosarcoma stem cells. miR-429-high and miR-429-low cells were separated using molecular probes and their stem cell-like properties were compared. The effect of miR-429 on osteosarcoma stem cell-like properties was further tested through transfection assays. Furthermore，the miR-429 downstream target gene was confirmed by luciferase assay.ResultsThe expression of miR-429 in osteosarcoma stem cells was much lower than that in control cells. miR-429-high cells displayed fewer stem celllike properties than did miR-429-low cells. These observations were confirmed by transfection assays. Additionally，our luciferase assays showed that miR-429 regulates Sox2 at the post-transcriptional level.ConclusionmiR-429 negatively regulates osteosarcoma stem celllike properties by targeting Sox2.

Keywordsosteosarcoma； stem cell；miR-429； Sox2

骨肉瘤是最常见的原发性恶性骨肿瘤，好发于青少年［1］。传统化疗药物具有全身毒性，且选择性差，约有10%～25%的患者会发生转移和耐药，治疗效果差［2］。因此，有效的骨肉瘤靶向治疗是临床上亟待解决的问题。

肿瘤干细胞是指能够自我更新和多向分化潜能的肿瘤细胞，在肿瘤的发生、转移和耐药中扮演重要角色［3］。GIBBS等［4］最先发现了骨肉瘤细胞里的干细胞，其能够形成具有自我更新能力的肿瘤球。ADHIKARI等［5］利用表面标记CD-117/Stro-1分选出骨肉瘤干细胞，且这群细胞具有显著增高的化疗耐药和远处转移特性。miRNAs是一类短链非编码RNA，并在转录水平上调节基因的表达。……