Expression of MicroRNA-301a and its Functional Roles in Malignant Melanoma
Expression of MicroRNA-301a and its Functional Roles in Malignant Melanoma
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DOI：
10.1159/000452540
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发表时间：
2016-01-01
影响因子：
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通讯作者：
Li, Xueyong
中科院分区：
文献类型：
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作者：
Cui, Lei;Li, Yuejun;Li, Xueyong
背景/目的：尽管microRNA-301a已被报道在许多人类癌症中作为癌基因发挥作用，但miR-301a在恶性黑色素瘤（MM）中的作用尚不清楚。本研究旨在探讨miR-301a在MM中的功能作用及其可能的分子机制。方法：采用实时荧光定量PCR （Quantitative real-time PCR, qRT-PCR）检测miR-301a在MM组织中的表达，分析其与MM患者转移及预后的相关性。在体外，通过过表达和敲低策略异位表达miR-301a，进一步研究miR-301a表达对MM细胞生长、凋亡、迁移、侵袭和化学敏感性的影响。此外，通过荧光素酶报告基因、qRT-PCR、Western blot检测鉴定了潜在和功能靶基因。结果：我们发现miR-301a在MM组织中表达明显上调，miR-301a的上调与MM患者的转移和预后不良相关。转染miR-301a/inhibitor可显著抑制MM细胞的生长、集落形成、迁移、侵袭，增强MM细胞的凋亡和化学敏感性，转染miR-301a/mimic可诱导MM细胞表型相反。荧光素酶报告基因、qRT-PCR和Western blot检测显示，磷酸酶和紧张素同源物（PTEN）是miR-301a的直接功能靶点。还观察到Aid和FAK信号通路参与了miR-301/ pten促进MM的进展。结论：综上所述，我们的研究表明mi R-301a可能被用作治疗人类MM的潜在治疗靶点。(C) 2016
Background/Aims: Although microRNA-301a has been reported to function as an oncogene in many human cancers, the roles of miR-301a in malignant melanoma (MM) is unclear. The present study aims to investigate the functional roles of miR-301a in MM and its possible molecular mechanisms. Methods: Quantitative real-time PCR (qRT-PCR) assay was performed to detect the expression of miR-301a in MM tissues, and analyze its correlation with metastasis and prognosis of MM patients. in vitro, mi R-301a was ectopically expressed using overexpression and knock-down strategies, and the effects of miR-301a expression on growth, apoptosis, migration, invasion and chemosensitivity of MM cells were further investigated. Furthermore, the potential and functional target gene was identified by luciferase reporter, qRT-PCR, Western blot assays. Results: We showed that the expression of miR-301a was significantly upregulated in MM tissues, and upregulation of miR-301a correlated with metastasis and poor prognosis of MM patients. Transfection of miR-301a/inhibitor significantly inhibited growth, colony formation, migration, invasion and enhanced apoptosis and chemosensitivity in MM cells, while transfection of miR-301a/mimic could induce the inverse effects on phenotypes of MM cells. Luciferase reporter, qRT-PCR and Western blot assays showed that phosphatase and tensin homolog (PTEN) was a direct and functional target of miR-301a. It was also observed that the Aid and FAK signaling pathways were involved in miR-301/PTEN-promoting MM progression. Conclusion: Taken together, our study suggests that mi R-301a may be used as a potential therapeutic target in the treatment of human MM. (C) 2016 The Author(s) Published by S. Karger AG, Basel