Li Zhi, Li Youjun, Wang Nan, Yang Lifeng, Zhao Wei, Zeng Xiandong
Department of Human Anatomy and Histoembryology, College of Basic Medical Sciences, Jilin University, China.
Department of Human Anatomy and Histoembryology, College of Basic Medical Sciences, Jilin University, China.
Biochem Biophys Res Commun. 2016 Mar 18;471(4):479-85. doi: 10.1016/j.bbrc.2016.02.050. Epub 2016 Feb 18.
miR-130b was significantly up-regulated in osteosarcoma (OS) cells. Naked cuticle homolog 2 (NKD2) inhibited tumor growth and metastasis in OS by suppressing Wnt signaling. We used three miRNA target analysis tools to identify potential targets of miR-130b, and found that NKD2 is a potential target of miR-130b. Based on these findings, we hypothesize that miR-130b might target NKD2 and regulate the Wnt signaling to promote OS growth. We detected the expression of miR-130b and NKD2 mRNA and protein by quantitative Real-Time PCR (qRT-PCR) and western blot assays, respectively, and found up-regulation of miR-130b and down-regulation of NKD2 mRNA and protein exist in OS cell lines. MTT and flow cytometry assays showed that miR-130b inhibitors inhibit proliferation and promote apoptosis in OS cells. Furthermore, we showed that NKD2 is a direct target of miR-130b, and miR-130b regulated proliferation and apoptosis of OS cells by targeting NKD2. We further investigated whether miR-130b and NKD2 regulate OS cell proliferation and apoptosis by inhibiting Wnt signaling, and the results confirmed our speculation that miR-130b targets NKD2 and regulates the Wnt signaling to promote proliferation and inhibit apoptosis of OS cells. These findings will offer new clues for OS development and progression, and novel potential therapeutic targets for OS.
miR-130b在骨肉瘤(OS)细胞中显著上调。裸角质同源物2(NKD2)通过抑制Wnt信号通路抑制OS中的肿瘤生长和转移。我们使用三种miRNA靶标分析工具来鉴定miR-130b的潜在靶标,发现NKD2是miR-130b的潜在靶标。基于这些发现,我们推测miR-130b可能靶向NKD2并调节Wnt信号通路以促进OS生长。我们分别通过定量实时PCR(qRT-PCR)和蛋白质印迹分析检测了miR-130b和NKD2 mRNA及蛋白的表达,发现在OS细胞系中存在miR-130b上调以及NKD2 mRNA和蛋白下调的情况。MTT和流式细胞术分析表明,miR-130b抑制剂可抑制OS细胞增殖并促进其凋亡。此外,我们表明NKD2是miR-130b的直接靶标,并且miR-130b通过靶向NKD2调节OS细胞的增殖和凋亡。我们进一步研究了miR-130b和NKD2是否通过抑制Wnt信号通路来调节OS细胞增殖和凋亡,结果证实了我们的推测,即miR-130b靶向NKD2并调节Wnt信号通路以促进OS细胞增殖并抑制其凋亡。这些发现将为OS的发生发展提供新线索,并为OS提供新的潜在治疗靶点。