微小RNA-203b-3p调控多发性骨髓瘤细胞增殖、迁移和侵袭的分子机制研究

Study on molecular mechanism of microRNA-203b-3p in regulating proliferation, migration and invasion of multiple myeloma cells

  • 摘要:
    目的 观察微小RNA-203b-3p (miR-203b-3p)通过调节肿瘤坏死因子超家族成员13b (TNFSF13B)对多发性骨髓瘤(MM)细胞的影响, 探讨miR-203b-3p抑制MM的相关机制。
    方法 采用实时荧光定量聚合酶链式反应(qRT-PCR)和蛋白质印迹分析(Western blot)检测miR-203b-3p和TNFSF13B在骨髓瘤肿瘤和细胞中的表达情况。miR-203b-3p与TNFSF13B的关系采用双荧光素酶报告实验进行验证。Lipofectamine 2000试剂用于MM细胞转染。miR-203b-3p和TNFSF13B对MM细胞生物功能的影响采用克隆形成试验、划痕试验和Transwell试验进行分析。
    结果 与癌旁组织及正常细胞比较, MM组织和细胞中miR-203b-3p的表达量相对较低,而TNFSF13B在MM肿瘤和细胞中的表达量与正常组织和细胞相比升高,差异有统计学意义(P < 0.05)。双荧光素酶报告实验结果表明, TNFSF13B是miR-203b-3p的直接靶点。miR-203b-3p在MM细胞中的过量表达能够抑制细胞的增殖和转移,差异有统计学意义(P < 0.05), 而TNFSF13B的表达上调则促进了MM细胞的增殖、迁移和侵袭,差异有统计学意义(P < 0.05)。
    结论 miR-203b-3p可能通过下调TNFSF13B抑制MM细胞的增殖、侵袭和转移。

     

    Abstract:
    Objective To observe the effect of microRNA-203b-3p (miR-203b-3p) on multiple myeloma (MM) cells by regulating tumor necrosis factor superfamily member 13b (TNFSF13B) and explore the related mechanism of miR-203b-3p in inhibiting MM.
    Methods Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) and Western blot were used to detect the expressions of miR-203b-3p and TNFSF13B in MM tumors and cells. The relationship between miR-203b-3p and TNFSF13B was validated by dual luciferase reporter experiments. Lipofectamine 2000 reagent was used for transfection of MM cells. The effects of miR-203b-3p and TNFSF13B on the biological function of MM cells were analyzed by colony formation assay, scratch assay, and Transwell assay.
    Results Compared with the cancer-adjacent tissues and normal cells, the expressions of miR-203b-3p in MM tissues and cells were relatively low, while the expression of TNFSF13B in MM tumors and cells was increased when compared with normal tissues and cells, and the differences were statistically significant (P < 0.05). The dual luciferase reporter experiment showed that TNFSF13B was the direct target of miR-203b-3p. Overexpression of miR-203b-3p in MM cells was able to inhibit cell proliferation and migration, while upregulation of TNFSF13B promoted MM cell proliferation, migration and invasion(P < 0.05).
    Conclusion MiR-203b-3p may inhibit the proliferation, invasion and migration of MM cells by downregulating TNFSF13B.

     

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