Wang R J, Shi K R, Zhang J, Zhang J, Gao R R, Zhu S C
Department of Human Anatomy and Embryology, Medical College of Tongji University, Shanghai 200092, China.
Zhonghua Bing Li Xue Za Zhi. 2016 Dec 8;45(12):866-870. doi: 10.3760/cma.j.issn.0529-5807.2016.12.010.
To investigate the effects of miR-93 on proliferation and apoptosis of osteosarcoma cells and the possible mechanism. The expression levels of miR-93 and the naked cuticle homolog 2 (NKD2) in 6 osteosarcoma cell lines (143B, HuO9, Saos2, MG63, U2OS and G292) and one osteoblast cell line hFOB1.19 were determined by quantitative real-time PCR and Western blot assays, respectively. MiR-93 down-regulated 143B and HuO9 cells were constructed by lipofection transfection, and their proliferation and apoptosis were detected by MTT and flow cytometry assays, respectively. Luciferase reporter assay was used to determine whether the 3'UTR of NKD2 mRNA was a binding target of miR-93. In addition, 143B cells were transfected with NKD2 cDNA, and the effects of NKD2 on proliferation and apoptosis of osteosarcoma cells were investigated. Up-regulation of miR-93 and down-regulation of NKD2 were detected in osteosarcoma cell lines. MTT and flow cytometry assays showed that miR-93 promoted proliferation and inhibited apoptosis in osteosarcoma cells. Luciferase assay confirmed that miR-93 targeted NKD2 directly. In addition, overexpression of NKD2 inhibited proliferation and promoted apoptosis of osteosarcoma cells were found. MiR-93 targets NKD2 to promote proliferation and inhibit apoptosis of osteosarcoma cells. The findings may have significant implications in the diagnosis and treatment of osteosarcoma.
探讨miR-93对骨肉瘤细胞增殖和凋亡的影响及其可能机制。分别采用定量实时PCR和蛋白质免疫印迹法检测6种骨肉瘤细胞系(143B、HuO9、Saos2、MG63、U2OS和G292)及1种成骨细胞系hFOB1.19中miR-93和无翅型表皮生长因子受体同源物2(NKD2)的表达水平。通过脂质体转染构建miR-93下调的143B和HuO9细胞,分别采用MTT法和流式细胞术检测其增殖和凋亡情况。采用荧光素酶报告基因检测法确定NKD2 mRNA的3'UTR是否为miR-93的结合靶点。此外,将NKD2 cDNA转染至143B细胞,研究NKD2对骨肉瘤细胞增殖和凋亡的影响。骨肉瘤细胞系中检测到miR-93上调和NKD2下调。MTT法和流式细胞术检测结果显示,miR-93促进骨肉瘤细胞增殖并抑制其凋亡。荧光素酶检测证实miR-93直接靶向NKD2。此外,还发现NKD2过表达抑制骨肉瘤细胞增殖并促进其凋亡。miR-93通过靶向NKD2促进骨肉瘤细胞增殖并抑制其凋亡。该研究结果可能对骨肉瘤的诊断和治疗具有重要意义。