Department of Otolaryngology Head and Neck Surgery, the Second Hospital, Jilin University, No. 218, Ziqiang Street, Nanguan District, Changchun, 130041, Jilin Province, China.
Department of Oral Geriatrics, School and Hospital of Stomatology, Jilin University, Changchun, 130021, China.
Cell Oncol (Dordr). 2020 Apr;43(2):249-261. doi: 10.1007/s13402-019-00485-5. Epub 2019 Dec 28.
The etiology of nasopharyngeal carcinoma (NPC) is multifactorial, complex and not fully characterized yet. MicroRNAs (miRNAs or miRs) have been found to contribute to the development and progression of NPC. Here, we aimed to investigate the putative role of miR-129-5p in NPC lymphangiogenesis and lymph node metastasis (LNM), including the involvement of its target gene ZIC2 and the Hedgehog signaling pathway.
The expression of miR-129-5p and ZIC2 in primary NPC tissues was assessed using RT-qPCR and Western blot analyses, followed by LNM and lymph vessel density (LVD) correlation analyses. A direct interaction between miR-129-5p and ZIC2 was verified using a dual-luciferase reporter assay. Gain- and loss-of-function experiments were conducted to investigate the effects of miR-129-5p and ZIC2 expression on NPC cell invasion, migration and proliferation in vitro, as well as on LDV and LNM in nude mice in vivo. Additionally, RT-qPCR and Western blot analyses were performed to determine the expression levels of Hedgehog signaling pathway-related factors.
We found that ZIC2 was highly expressed, and miR-129-5p was lowly expressed, in primary NPC tissues. In addition, we found that miR-129-5p can directly bind to and reduce ZIC2 expression. LVD was found to be negatively correlated with miR-129-5p and to be positively correlated with ZIC2 expression. Concomitantly, we found that miR-129-5p abrogated activation of the Hedgehog signaling pathway via ZIC2 targeting, leading to suppression of NPC cell invasion, migration and proliferation in vitro as well as suppression of LNM and LVD in vivo.
From our data we conclude that miR-129-5p, by decreasing ZIC2 expression, may inhibit NPC lymphangiogenesis and LNM through suppression of the Hedgehog signaling pathway.
鼻咽癌（NPC）的病因是多因素的、复杂的，尚未完全阐明。现已发现 microRNAs（miRNAs 或 miRs）有助于 NPC 的发展和进展。在这里，我们旨在研究 miR-129-5p 在 NPC 淋巴管生成和淋巴结转移（LNM）中的潜在作用，包括其靶基因 ZIC2 和 Hedgehog 信号通路的参与。
使用 RT-qPCR 和 Western blot 分析评估原发性 NPC 组织中 miR-129-5p 和 ZIC2 的表达情况，然后进行 LNM 和淋巴管密度（LVD）相关性分析。使用双荧光素酶报告基因测定验证 miR-129-5p 和 ZIC2 之间的直接相互作用。进行增益和失能实验，以研究 miR-129-5p 和 ZIC2 表达对 NPC 细胞体外侵袭、迁移和增殖的影响，以及体内裸鼠 LVD 和 LNM 的影响。此外，进行 RT-qPCR 和 Western blot 分析以确定 Hedgehog 信号通路相关因子的表达水平。
我们发现 ZIC2 在原发性 NPC 组织中高表达，miR-129-5p 低表达。此外，我们发现 miR-129-5p 可以直接结合并降低 ZIC2 的表达。LVD 与 miR-129-5p 呈负相关，与 ZIC2 的表达呈正相关。同时，我们发现 miR-129-5p 通过靶向 ZIC2 使 Hedgehog 信号通路失活，从而抑制 NPC 细胞在体外的侵袭、迁移和增殖，并抑制体内 LNM 和 LVD。
根据我们的数据，我们得出结论，miR-129-5p 通过降低 ZIC2 的表达，可能通过抑制 Hedgehog 信号通路来抑制 NPC 的淋巴管生成和 LNM。
