MicroR-760 suppresses cancer stem cell subpopulation and breast cancer cell proliferation and metastasis: By down-regulating NANOG
MicroR-760 suppresses cancer stem cell subpopulation and breast cancer cell proliferation and metastasis: By down-regulating NANOG
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DOI：
10.1016/j.biopha.2016.03.024
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发表时间：
2016-05-01
影响因子：
7.5
通讯作者：
Wang, Yi-meng
中科院分区：
文献类型：
--
作者：
Han, Ming-li;Wang, Fang;Wang, Yi-meng
背景和目的：新的证据表明，肿瘤干细胞与肿瘤的侵袭、转移和治疗耐药有关。目前，乳腺癌干细胞（BCSC）群体在肿瘤转移中的作用机制尚不清楚。方法：采用流式细胞仪检测乳腺癌MCF-7和BT-549细胞中BCSC的表面标志物（CD 44（+）/CD 24（-/low）），并分析其表达情况。使用定量RT-PCR评估miR-760和NANOG mRNA表达。使用蛋白质印迹法测定NANOG蛋白的表达。MTT法检测细胞增殖;结果：BT-549细胞的CD 44（+）/CD 24（-/low）亚群明显多于MCF-7细胞。BT-549细胞中miR-760的表达水平低于MCF-7细胞，NANOG的表达水平高于MCF-7细胞。通过对细胞miR-760调控的结果，我们发现miR-760过表达抑制了CD 44（+）/CD 24（-/low）群体，并抑制了BT-549的细胞增殖和迁移。相反，miR-760的敲低促进了MCF-7细胞的CD 44（+）/CD 24（-/low）群体和迁移。通过荧光素酶报告基因分析，证实miR-760通过调控NANOG的表达与NANOG功能相关。结论：以miR-760/NANOG轴为靶点，可能成为抑制乳腺癌干细胞亚群从而预防肿瘤转移的新途径。(C)2016年由Elsevier Masson SAS出版。
Background and objective: Emerging evidences suggest that cancer stem cells are responsible for tumor aggressive, metastasis and therapeutic resistance. To data, the mechanism underlying breast cancer stem cell (BCSC) population within tumor metastasis remains to be fully elucidated. The current study was to investigate the potential role of microRNA-760 (miR-760) and its associated target gene in population and metastasis of BCSC.Methods: Characteristic BCSCs surface markers (CD44(+)/CD24(-/low)) were determined by flow cytometry in breast cancer MCF-7 and BT-549 cells. Quantitative RT-PCR was used to evaluate miR-760 and NANOG mRNA expression. Expression of NANOG protein was determined using western blot. Cell proliferation was determined by MTT assay. The model of breast cancer cell xenograft was used to evaluate the effect of miR-760 on tumor growth.Results: BT-549 cell has substantially more CD44(+)/CD24(-/low) subpopulation than MCF-7 cell. Moreover, BT-549 cell expressed lower level of miR-760 and higher level of NANOG than MCF-7cell. By result from cellular miR-760 modulation, we found that miR-760 overexpression suppressed CD44(+)/CD24(-/low) population as well as inhibited cell proliferation and migration of BT-549. On the contrary, knockdown of miR-760 promoted CD44(+)/CD24(-/low) population and migration of MCF-7 cells. By luciferase reporter assay, miR-760 was proved to be functional associated with NANOG via regulating its expression. This functional interaction was showed to be involved in controlling proliferation and migration of MCF-7 and BT-549 cell.Conclusion: These data suggest that the target of miR-760/NANOG axis may represent a new therapeutic approach to suppress breast cancer stem cell subpopulation thereby prevent cancer metastasis. (C) 2016 Published by Elsevier Masson SAS.