MiR-483-3p, Mediated by KLF9, Functions as Tumor Suppressor in Testicular Seminoma via Targeting MMP9.

Lei Zhang,Wei Wang,Shenglin Gao,Kai Xu,Li Zuo,Zhiqiang Qin,Yashi Ruan,Xian Gao,Shouyong Liu,Xiaokai Shi,Wei Zhang,Lifeng Zhang,Kai Zhu

doi:10.3389/fonc.2020.596574

Lei Zhang, Wei Wang + Show 11 more

Open Access

https://doi.org/10.3389/fonc.2020.596574

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Abstract

BackgroundSeminoma (SEM) is the most frequent testicular germ cell tumor with a high incidence in young men. The present study aims to explore the function and regulatory mechanism of miR-483-3p in SEM.MethodsRT-qPCR was performed to investigate miR-483-3p levels in SEM tissues. The effect of miR-483-3p on TCam-2 cells was assessed by CCK-8, colony formation, cell migration, and invasion assays. Luciferase reporter assays were performed to investigate the interaction between miR-483-3p and MMP9, and then the recovery experiments were performed. Moreover, the potential upstream regulator of miR-483-3p was predicted based on JASPAR database.ResultsmiR-483-3p was down-regulated in SEM tissues versus paracancerous normal tissues. The expression level of miR-483-3p was significantly associated with tumor stage by RT-qPCR. Functionally, miR-483-3p over-expression suppressed cell growth, migration, and invasion in SEM cell lines. Mechanically, miR-483-3p negatively regulated MMP9 by directly binding to its 3′-UTR. The over-expression of miR-483-3p could reverse the promoting role of MMP9 over-expression on the proliferation, migration, and invasion of TCam-2 cells. Moreover, KLF9 was identified as a potential upstream regulator of miR-483-3p and functions as a tumor suppressor.ConclusionsIn general, our study suggested that miR-483-3p could inhibit the cell growth, migration, and invasion of testicular SEM by targeting MMP9. Moreover, KLF9 is an upstream positive regulator of miR-483-3p and also functions as a tumor suppressor in SEM.

Highlights

Testicular germ cell tumor (TGCT) is the most common solid malignancy in young men with the incidence rising constantly over the past decades [1, 2]
We performed RT-qPCR to detect the mRNA level of miR-483-3p in 20 pairs of SEM and paracancerous tissues from patients with testicular SEM
TCam-2 cells were transfected with lentiviral constructs to the over-express or inhibit the expression of miR-483-3p, and the transfection efficiencies were validated by RT-qPCR (Figure 2A)

Summary

Introduction

Testicular germ cell tumor (TGCT) is the most common solid malignancy in young men with the incidence rising constantly over the past decades [1, 2]. The majority (~70%) of SEM patients present with clinical stage I disease and have a very good overall survival after surgical treatment and radiotherapy [6]. Higher-dose radiotherapy and platin-based chemotherapy are recommended for Stage II/ III SEM with lymph nodes and distant metastasis, which reveals high risks and unfavorable prognosis [6]. It is imperative to further illuminate the molecular mechanisms involved in the progression of SEM and provide potential biomarkers for the clinical treatment, surveillance, and prognosis of patients with SEM. Seminoma (SEM) is the most frequent testicular germ cell tumor with a high incidence in young men. The present study aims to explore the function and regulatory mechanism of miR-483-3p in SEM

Methods

Results

Conclusion