Liu Wei, Liu Sheng-Yao, He Yong-Bin, Huang Rui-Liang, Deng Song-Yun, Ni Guo-Xin, Yu Bin
Department of Orthopeadics and Traumatology, Nanfang Hospital, Southern Medical University, 1838 Guangzhou Avenue (N), Guangzhou 510515, China; Department of Orthopedics, The People's Hospital of Gaoming District of Foshan City, Guangdong 528500, China.
Department of Orthopeadics and Traumatology, Nanfang Hospital, Southern Medical University, 1838 Guangzhou Avenue (N), Guangzhou 510515, China.
Biomed Pharmacother. 2017 Mar;87:621-627. doi: 10.1016/j.biopha.2016.12.121. Epub 2017 Jan 10.
Previous studies have shown that MiR-451 plays an important role in human osteosarcoma carcinogenesis, but the underlying mechanism by which MiR-451 affects the osteosarcoma has not been fully understood. This study intends to uncover the mechanism by which MiR-451 functions as a tumor suppressor. The expression of MiR-451 in osteosarcoma tissues and osteosarcoma cell lines was monitored by real-time PCR. The proliferation ability was examined by MTT and cell cycle assay. The migration and apoptosis of cells were monitored by migration assay and flow cytometry, respectively. Moreover, the angiogenesis of HUVEC cells transfected with MiR-451 mimics was examined by tube formation assay. The effect of MiR-451 on MIF was determined by luciferase assays and Western blot assay. The results showed that MiR-451 expression level was significantly reduced in the osteosarcoma compared with normal bone tissues. Overexpression of MiR-451 significantly attenuated the proliferation and migration, and induced the apoptosis of osteosarcoma cells. Furthermore, the angiogenesis of HUVEC cells transfected with MiR-451 mimics was assayed and the decreased angiogenic ability was detected compared to the controls. Finally, we demonstrated that MiR-451 overexpression inhibited the malignant behavior of osteosarcoma by downregulating MIF. These findings suggest that MiR-451 may act as a tumor suppressor in osteosarcoma. MiR-451 inhibited cell proliferation, migration and angiogenesis and promoted apoptosis of human osteosarcoma cells, at least partially, by inhibiting the expression of MIF. MiR-451/MIF may be a novel therapeutic target in treatment of osteosarcoma.
先前的研究表明，MiR-451在人类骨肉瘤致癌过程中发挥重要作用，但MiR-451影响骨肉瘤的潜在机制尚未完全明确。本研究旨在揭示MiR-451作为肿瘤抑制因子发挥作用的机制。通过实时PCR监测MiR-451在骨肉瘤组织和骨肉瘤细胞系中的表达。采用MTT法和细胞周期分析法检测细胞增殖能力。分别通过迁移试验和流式细胞术监测细胞的迁移和凋亡。此外，通过管腔形成试验检测转染MiR-451模拟物的人脐静脉内皮细胞（HUVEC）的血管生成情况。通过荧光素酶报告基因检测和蛋白质免疫印迹法确定MiR-451对巨噬细胞移动抑制因子（MIF）的影响。结果显示，与正常骨组织相比，MiR-451在骨肉瘤中的表达水平显著降低。MiR-451的过表达显著减弱了骨肉瘤细胞的增殖和迁移，并诱导其凋亡。此外，对转染MiR-451模拟物的HUVEC细胞进行血管生成检测，发现与对照组相比其血管生成能力下降。最后，我们证明MiR-451的过表达通过下调MIF抑制骨肉瘤的恶性行为。这些发现表明，MiR-451可能在骨肉瘤中作为肿瘤抑制因子发挥作用。MiR-451至少部分地通过抑制MIF的表达来抑制人骨肉瘤细胞的增殖、迁移和血管生成，并促进其凋亡。MiR-451/MIF可能是骨肉瘤治疗中的一个新的治疗靶点。
