Department of Hematology, Hanzhong Central Hospital, Hanzhong, 723000, China.
Department of Hematology, the First Affiliated Hospital of Xi'an Jiao Tong University, Xi'an, 710061, China.
Cell Biol Int. 2019 Nov;43(11):1223-1233. doi: 10.1002/cbin.11104. Epub 2019 Jul 17.
miR-9 has been reported to play a pivotal role in multiple human cancers by acting as an oncogene or tumor suppressor. In this study, we explored the possible role and molecular mechanism of miR-9 in multiple myeloma (MM). The miR-9 expression was examined by quantitative real-time polymerase chain reaction assay. Transfection with miR-9-mimics, miR-9-inhibitor, pcDNA-TRIM56, or si-TRIM56 into cells was used to change the expression levels of miR-9 and TRIM56. Western blot analysis was used to detect the expression of TRIM56, p65, p-p65, IκBα, and p-IκBα. The potential target of miR-9 was confirmed by luciferase reporter assay. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay, colony formation assay, and flow cytometry were used to assess the abilities of cell proliferation and apoptosis. miR-9 was upregulated in MM patients and cell lines, and miR-9 overexpression promoted proliferation and repressed apoptosis in MM cell lines. TRIM56 was confirmed as a target of miR-9. Moreover, TRIM56 reversed miR-9-mediated pro-proliferation and anti-apoptosis effect on MM cell lines. Furthermore, nuclear factor-κB (NF-κB) pathway was involved in miR-9/TRIM56-mediated regulation on MM cell lines. miR-9 promoted the development and progression of MM by regulating TRIM56/NF-κB pathway, thereby providing a potential microRNA-based target for MM therapy.
miR-9 在多种人类癌症中通过作为癌基因或肿瘤抑制因子发挥关键作用。在这项研究中，我们探讨了 miR-9 在多发性骨髓瘤（MM）中的可能作用和分子机制。通过定量实时聚合酶链反应（PCR）检测 miR-9 的表达。通过转染 miR-9 模拟物、miR-9 抑制剂、pcDNA-TRIM56 或 si-TRIM56 进入细胞来改变 miR-9 和 TRIM56 的表达水平。Western blot 分析用于检测 TRIM56、p65、p-p65、IκBα 和 p-IκBα 的表达。通过荧光素酶报告基因检测证实了 miR-9 的潜在靶标。3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑（MTT）测定、集落形成测定和流式细胞术用于评估细胞增殖和凋亡的能力。miR-9 在 MM 患者和细胞系中上调，miR-9 过表达促进 MM 细胞系的增殖并抑制凋亡。TRIM56 被确认为 miR-9 的靶标。此外，TRIM56 逆转了 miR-9 对 MM 细胞系的促增殖和抗凋亡作用。此外，核因子-κB（NF-κB）途径参与了 miR-9/TRIM56 对 MM 细胞系的调节。miR-9 通过调节 TRIM56/NF-κB 通路促进 MM 的发生和发展，从而为 MM 治疗提供了一种潜在的基于 microRNA 的靶标。
