Department of Orthopaedics, Rongcheng People's Hospital of Shandong Province , Rongcheng, Shandong, China.
Department of Neurology, Rongcheng People's Hospital of Shandong Province , Rongcheng, Shandong, China.
Cancer Biol Ther. 2020 Nov 1;21(11):1051-1059. doi: 10.1080/15384047.2020.1831373.
Multiple functions of miR-199b-5p in diseases have been demonstrated by existing studies. However, never has the correlation between miR-199b-5p and multiple myeloma (MM) been established.
qRT-PCR analyzed RNA expression and western blot measured protein expression. Cell proliferation ability was tested via colony formation and EdU assays, and apoptosis was determined via TUNEL, flow cytometry and detection of apoptosis-related proteins. Position of LRRC75A antisense RNA 1 (LRRC75A-AS1) was recognized by FISH assay. RIP, RNA pull-down and luciferase reporter experiments explored the molecular interplay.
GEO (Gene Expression Omnibus) data revealed miR-199b-5p upregulation in MM specimens, and qRT-PCR data verified miR-199b-5p upregulation in MM cells. Inhibiting miR-199b-5p markedly impeded MM cell proliferation and stimulated apoptosis. Moreover, we demonstrated the mechanism that miR-199b-5p was decoyed by LRRC75A-AS1 and miR-199b-5p targeted programmed cell death 4 (PDCD4) to repress its expression. Further, LRRC75A-AS1 was verified to repress proliferation and prompt apoptosis in a PDCD4-dependent way in MM cells.
Our data displayed that miR-199b-5p was sequestered by LRRC75A-AS1 so that PDCD4 was released to repress MM, implying the targeting miR-199b-5p as a novel thought for improving MM therapy.
现有研究已经证实了 miR-199b-5p 在多种疾病中的多种功能。然而，miR-199b-5p 与多发性骨髓瘤（MM）之间的相关性从未得到证实。
qRT-PCR 分析 RNA 表达，Western blot 检测蛋白表达。通过集落形成和 EdU 测定试验检测细胞增殖能力，通过 TUNEL、流式细胞术和检测凋亡相关蛋白来确定细胞凋亡。通过 FISH 检测识别 LRRC75A 反义 RNA 1（LRRC75A-AS1）的位置。RIP、RNA 下拉和荧光素酶报告基因实验探讨了分子相互作用。
GEO（基因表达综合数据库）数据显示 MM 标本中 miR-199b-5p 上调，qRT-PCR 数据验证了 MM 细胞中 miR-199b-5p 的上调。抑制 miR-199b-5p 显著抑制 MM 细胞增殖并刺激细胞凋亡。此外，我们证明了 miR-199b-5p 被 LRRC75A-AS1 捕获，miR-199b-5p 靶向程序性细胞死亡因子 4（PDCD4）抑制其表达的机制。此外，LRRC75A-AS1 被证实以 PDCD4 依赖的方式在 MM 细胞中抑制增殖并促进凋亡。
我们的数据显示，miR-199b-5p 被 LRRC75A-AS1 封闭，从而释放 PDCD4 抑制 MM，表明靶向 miR-199b-5p 可能是改善 MM 治疗的新策略。
