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张怀斌, 杨国清, 梁文强, 谢犇, 王勇平. MiR-129-5p靶向HMGB1抑制骨肉瘤细胞增殖和迁移[J]. 肿瘤防治研究, 2022, 49(1): 5-11. DOI: 10.3971/j.issn.1000-8578.2022.21.0707
引用本文: 张怀斌, 杨国清, 梁文强, 谢犇, 王勇平. MiR-129-5p靶向HMGB1抑制骨肉瘤细胞增殖和迁移[J]. 肿瘤防治研究, 2022, 49(1): 5-11. DOI: 10.3971/j.issn.1000-8578.2022.21.0707
ZHANG Huaibin, YANG Guoqing, LIANG Wenqiang, XIE Ben, WANG Yongping. MiRNA-129-5p Targeting HMGB1 Inhibits Proliferation and Migration of Osteosarcoma Cells[J]. Cancer Research on Prevention and Treatment, 2022, 49(1): 5-11. DOI: 10.3971/j.issn.1000-8578.2022.21.0707
Citation: ZHANG Huaibin, YANG Guoqing, LIANG Wenqiang, XIE Ben, WANG Yongping. MiRNA-129-5p Targeting HMGB1 Inhibits Proliferation and Migration of Osteosarcoma Cells[J]. Cancer Research on Prevention and Treatment, 2022, 49(1): 5-11. DOI: 10.3971/j.issn.1000-8578.2022.21.0707

MiR-129-5p靶向HMGB1抑制骨肉瘤细胞增殖和迁移

MiRNA-129-5p Targeting HMGB1 Inhibits Proliferation and Migration of Osteosarcoma Cells

  • 摘要:
    目的 探讨miR-129-5p对骨肉瘤(OS)细胞增殖和迁移的影响以及对HMGB1的调控作用。
    方法 RT-PCR和Western blot法分别检测骨肉瘤细胞株MG-63、Saos-2和成骨细胞hFOB1.19中miR-129-5p和HMGB1的表达。生物信息学预测miR-129-5p与HMGB1基因是否存在结合位点,双荧光素酶报告基因检测验证miR-129-5p和HMGB1的相互作用。将miR-129-5p模拟物和抑制物分别转染到miR-129-5p表达量较低和较高的骨肉瘤细胞株中,RT-PCR检测转染效率。转染成功后分别用CCK-8法、划痕实验和Transwell迁移实验检测OS细胞株的增殖和迁移能力;Western blot检测转染后OS细胞株中HMGB1的表达。
    结果 miR-129-5p在OS细胞中的表达低于正常成骨细胞(P < 0.05),HMGB1在OS细胞株中的表达高于正常成骨细胞(P < 0.05)。miR-129-5p和HMGB1基因存在结合位点,miR-129-5p mimic转染能下调HMGB1-WT质粒组的荧光素酶活性(P < 0.05)。转染miR-129-5p模拟物可显著提高MG-63细胞中miR-129-5p的表达量(P < 0.05),且可抑制MG-63细胞的增殖和迁移(P < 0.05),同时降低HMGB1的表达水平;转染miR-129-5p抑制物后可显著降低Saos-2细胞中miR-129-5p的表达量(P < 0.05),且可增强Saos-2细胞增殖和迁移能力(P < 0.05),同时提高HMGB1表达水平。
    结论 miR-129-5p可能通过HMGB1基因抑制骨肉瘤细胞增殖和迁移。

     

    Abstract:
    Objective To investigate the effects of miR-129-5p on the proliferation and migration of osteosarcoma cells and the regulation of HMGB1 gene.
    Methods The expression of miR-129-5p and HMGB1 in osteosarcoma cell line MG-63, Saos-2 and osteoblast hFOB1.19 were detected by RT-PCR and Western blot. Bioinformatics methods were used to predict whether there were binding sites between mir-129-5p and HMGB1 gene. Double luciferase reporter gene system was used to verify the interaction between miR-129-5p and the target gene HMGB1. miR-129-5p mimic and inhibitor were transfected into osteosarcoma cell lines with low and high miR-129-5p expression, respectively, and the transfection efficiency was detected by RT-PCR. After successful transfection, the proliferation and migration of osteosarcoma cell lines were detected by CCK-8 assay, scratch assay and Transwell migration assay, respectively, and Western blot was used to detect the expression of HMGB1 in the transfected osteosarcoma cell lines.
    Results Expression of miR-129-5p in osteosarcoma cells was lower than that in normal osteoblasts (P < 0.05), and the expression of HMGB1 in osteosarcoma cell lines was higher than that in normal osteoblasts (P < 0.05). There were binding sites between miR-129-5p and HMGB1 genes, and the luciferase activity of HMGB1-WT plasmid group was down-regulated after transfection with miR-129-5p mimic (P < 0.05). Transfection of miR-129-5p mimic significantly increased the expression of miR-129-5p in MG-63 cells (P < 0.05), inhibited the proliferation and migration of MG-63 cells (P < 0.05), and decreased the expression level of HMGB1. After transfection with miR-129-5p inhibitor, the expression of miR-129-5p in Saos-2 cells was significantly decreased (P < 0.05), the proliferation and migration abilities of Saos-2 cells were enhanced (P < 0.05), and the expression level of HMGB1 was also increased.
    Conclusion miR-129-5p may inhibit the proliferation and migration of osteosarcoma cells through HMGB1 gene.

     

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