miR-106b-5p promotes cell cycle progression of malignant melanoma by targeting PTEN
miR-106b-5p promotes cell cycle progression of malignant melanoma by targeting PTEN
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DOI：
10.3892/or.2017.6099
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发表时间：
2018-01-01
期刊：
影响因子：
4.2
通讯作者：
Sheng, Liang
中科院分区：
文献类型：
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作者：
Chen, Xu-E;Chen, Pu;Sheng, Liang
本研究探讨miR-106b-5p/PTEN信号转导通路对恶性黑色素瘤细胞周期的影响。用qRT-PCR方法检测MIR-106b-5p基因的表达。通过瞬时转染法，miR-106b-5p或PTEN在MM细胞中表达上调或下调。采用四甲基偶氮唑盐比色法、集落形成实验和流式细胞术检测miR-106b-5p和PTEN对细胞周期的影响。Western印迹分析定量所有蛋白，荧光素酶报告实验验证针对PTEN的miR-106b-5p。MIR-106b-5p在MM组织和细胞系中过表达。上调miR-106b-5p的MM细胞生长较快，细胞周期较短，而miR-106b-5p下调的MM细胞表现出相反的趋势。PTEN受miR-106b-5p转录后调控。在此基础上，进一步研究了细胞周期蛋白D1和P27(Kip1)之间的相互作用，发现miR-106b-5p可通过Akt/ERK途径刺激细胞周期蛋白D1，抑制P27(Kip1)。本研究结果提示miR-106b-5p可能通过靶向PTEN从而调控下游细胞周期相关蛋白和Akt/ERK通路，在多发性骨髓瘤的发生发展中起到启动子的作用。
This study investigated how miR-106b-5p/PTEN signaling affects the cell cycle of malignant melanoma (MM) cells. miR-106b-5p mRNA was identified with qRT-PCR. Through transient transfection, miR-106b-5p or PTEN was upregulated and downregulated in MM cells. With such transfected cells, MTT assay, colony formation assay and flow cytometry were carried out to investigate the role of miR-106b-5p in cell cycle progression after the transfected cells were treated with reverse-regulation of miR-106b-5p or PTEN. Western blot analysis was used to quantify all proteins, and a luciferase reporter assay was carried out to validate miR-106b-5p targeting PTEN. miR-106b-5p mRNA was overexpressed in MM tissues and cell lines. MM cells with upregulated miR-106b-5p presented faster growth and shorter cell cycles, while those with knockdown of miR-106b-5p presented the opposite trend. PTEN was subject to post-transcriptional regulation of miR-106b-5p. Based on such a finding, further exploration was carried out to investigate the interaction between cyclin D1 and P27(Kip1), with the finding that miR-106b-5p can stimulate cyclin D1 and suppress P27(Kip1) via the Akt/ERK pathway. The results of this study suggest that miR-106b-5p may be a promoter in MM progression, possibly by targeting PTEN and thus regulating the downstream cell-cycle-related proteins and Akt/ERK pathway.