Elsevier

Life Sciences

Volume 175, 15 April 2017, Pages 47-51
Life Sciences

miR-519 suppresses nasopharyngeal carcinoma cell proliferation by targeting oncogene URG4/URGCP

https://doi.org/10.1016/j.lfs.2017.03.010Get rights and content

Abstract

Nasopharyngeal carcinoma (NPC) is one of the most common malignant tumor in Southeast Asia, its regulatory mechanism is still to be understood. miR-519 inhibits the progression of several tumors, including cervical cancer, ovarian cancer and gastric cancer. But its role in NPC hasn't been studied. In present study, we found miR-519 was downregulated in NPC cells, its overexpression inhibited NPC cell proliferation and arrested cell cycle at G0/G1 phase, while its knockdown promoted NPC cell proliferation and cell cycle progression. An oncogene URG4/URGCP (upregulated gene-4/upregulator of cell proliferation) was the target of miR-519, URG4 was upregulated in NPC cells, miR-519 inhibited URG4 expression by directly binding to the 3′UTR of URG4. miR-519 inhibited Cyclin D1 expression and the phosphorylation level of Rb, and increased p21 and p27 expression, confirming miR-519 blocked G1/S transition. Moreover, miR-519 level was negative correlated with URG4 level in NPC tissues. In summary, we found miR-519 NPC cell proliferation by inhibiting URG4.

Introduction

Nasopharyngeal carcinoma (NPC) is an endemic non-lymphomatous squamous-cell malignant tumor, people lived in southern China have the highest incidence, Epstein-Barr virus is the main inducing factor for NPC, other factors also have been shown to regulate PNC initiation and development, but the detailed molecular regulatory mechanisms are still to be understood [12], [19].

microRNA (miRNA) is a small non-coding RNA, it regulates gene expression through degrading target mRNA or/and inhibiting mRNA translation. It could serve as the prognostic factor or therapy target for NPC [6], [11], [17]. For example, miR-9 inhibits NPC growth and metastasis by inhibiting chemokine receptor CXCR4, interferon-regulated genes and MHC class I molecules, plasma miR-9 might be a biomarker for NPC metastasis [5], [8], [9]. miR-214 promotes NPC cell proliferation, invasion and tumorigenesis through inhibiting lactotransferrin (LTF) to activate AKT pathway [3]. miR-216b inhibits NPC growth and invasion by inhibiting KRAS, in turn inactivating AKT and ERKs pathways [2]. Previous reports show miR-519 can acts as tumor suppressor, miR-519 inhibits the proliferation of several cancer cells by targeting RNA-binding protein HuR, cervical cancer, ovarian cancer, gastric cancer are included [1], [7], [10], [20], miR-519 inhibits colorectal cancer cell proliferation by inhibiting Orail, in turn activating AKT/GSK3β pathway [4]. But the role of miR-519 in NPC cell proliferation hasn't been reported. Here, we studied the role of miR-519 in NPC cell proliferation, and found miR-519 inhibits NPC cell proliferation by inhibiting URG4.

Section snippets

Cells and specimens

Normal nasopharyngeal epithelium cell (NPEC) was maintained in Keratinocyte-SFM medium (Invitrogen) supplemented with bovine pituitary extract. NPC cells CNE1, CNE2, HONE1, SUNE1, C666 and 6-10B were maintained in RPMI-1640 supplemented with 10% fetal bovine serum (FBS, Hyclone). Eight pairs of primary NPC biopsies and matched adjacent normal nasopharyngeal epithelium biopsies were collected from Affiliated Hospital of Guizhou Medical University and Guizhou Cancer Hospital. Informed consent was

miR-519 overexpression inhibits NPC cells proliferation

To examine whether miR-519 regulates NPC development, miR-519 levels in NPC cells and normal nasopharyngeal epithelium cell (NPEC) were analyzed, it was significantly downregulated in NPC cells (Fig. 1A). We transfected miR-519 mimic into NPC cells SUNE1 and CNE1, and found miR-519 was significantly upregulated in NPC cells transfected with miR-519 mimic compared to the negative control (NC) (Fig. 1B). Colony formation assay and MTT assay suggested miR-519 overexpression significantly inhibited

Discussion

In present study, we found miR-519 inhibited NPC cell proliferation, and arrested cell cycle at G0/G1 phase. Mechanism analysis suggested URG4 was the target of miR-519, miR-519 directly bound to the 3′UTR of URG4 determined by luciferase reporter system. To confirm miR-519 blocks G1/S transition, we evaluated the effect of miR-519 on Cyclin D1, p21, p27, Rb and p-Rb, qRT-PCR and western blot assay suggested miR-519 overexpression inhibited Cyclin D1 expression and the phosphorylation level of

Conflict of interest

None.

Acknowledgments

This study was supported by the Foundation of Guizhou Science and Technology Department (No. TN2014-53, No. D2011-16 and No. E2006-12), the Foundation of Governor of Guizhou Province (No. S2010-8), and the Foundation of Guizhou Provincial Education Department (No. F2005-8 and No. F2010-8).

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