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• W2907172009 abstract "To investigate the microRNA (miR)-150 expression level in human osteosarcoma cell lines (Saos-2, MG-63) and its function in cell proliferation, and to explore the potential molecular mechanisms. Methods: MiR-150 expression levels in human osteosarcoma cell lines (Saos-2, MG-63) and normal osteoblast cell line (NHOst) were detected by relative quantitative real-time PCR (qRT-PCR). MiR-150 was overexpressed in Saos-2 and MG-63 cells by lentivirus infection. Cell proliferation rates were monitored by MTS assay. RUNX2 and β-actin protein levels were examined by Western blot. Inhibitory effect of miR-150 on binding RUNX2 3'-UTR was detected by Dual-Luciferase assay. Results: MiR-150 expression level is lower in human osteosarcoma cell lines (Saos-2, MG-63) compared to the normal osteoblast cell line (NHOst) (0.23±0.02 and 0.32±0.03 vs 1.00±0.02), which showed statistical significance (P<0.01). After lentivirus infection, miR-150 level increased in Saos-2 (P<0.01) and MG-63 cells (P<0.01). Overexpression of miR-150 decreased cell proliferation and RUNX2 protein level in Saos-2 and MG-63 cells. The binding of miR-150 to RUNX2 3'-UTR decreased luciferase activity by 69% in Saos-2 cells (P<0.05) and 59% in MG-63 cells (P<0.05). Administration of exogenous RUNX2 recovered the cell proliferation in miR-150 overexpressed Saos-2 and MG-63 cell lines (P<0.01). Conclusion: MiR-150 inhibites proliferation in human osteosarcoma cell lines through binding to RUNX2 3'-UTR, resulting in the reducion of RUNX2 protein level.目的：研究人骨肉瘤细胞系(Saos-2，MG-63)中microRNA-150(miR-150)表达水平及其对细胞增殖的作用，并探讨其分子生物学作用机制。方法：采用相对实时定量PCR检测人骨肉瘤细胞系Saos-2，MG-63和正常成骨细胞系NHOst中miR-150表达差异。采用慢病毒转染过表达miR-150，实时定量PCR检测过表达组miR-150水平。细胞增殖实验采用MTS[3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4- sulfophenyl)- 2H-tetrazolium] 法。Western印迹检测Runt相关转录因子2(Runt-related transcription factor 2，RUNX2)及内参β-actin蛋白水平。双荧光素酶反应检测miR-150与RUNX2 3'-UTR结合对上游基因表达抑制率。结果：MiR-150在Saos-2和MG-63细胞系中表达低于正常成骨细胞系NHOst，分别为0.23±0.02，0.32±0.03和1.00±0.02，差异具有统计学意义(P<0.01)。Saos-2细胞系空载体对照组和miR-150过表达组分别为1.00±0.08和4.17±0.19，差异具有统计学意义(P<0.01)，MG-63细胞系空载体对照组和miR-150过表达组分别为1.00±0.07和3.43±0.10，差异具有统计学意义(P<0.01)。过表达miR-150后，Saos-2和MG-63细胞增殖率明显下降(P<0.01)，RUNX2蛋白量明显降低(P<0.01)。Saos-2细胞系中， miR-150结合RUNX2 3'-UTR区抑制69%上游荧光素酶活性(P<0.05)；MG-63细胞系中，miR-150抑制59%上游荧光素酶活性(P<0.05)。在过表达miR-150的Saos-2和MG-63细胞中，补充外源性RUNX2蛋白，可恢复骨肉瘤细胞的增殖水平(P<0.01)。结论：MiR-150具有抑制骨肉瘤细胞增殖的作用，直接结合RUNX2 3'-UTR抑制转录因子RUNX2合成是其重要的作用机制。." @default.
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• W2907172009 date "2016-12-28" @default.
• W2907172009 modified "2023-09-26" @default.
• W2907172009 title "[MicroRNA-150 inhibits osteosarcoma cell proliferation by targeting RUNX2 gene]." @default.
• W2907172009 doi "https://doi.org/10.11817/j.issn.1672-7347.2016.12.006" @default.
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