Lei Zhang1,2, Yashi Ruan2,3,4, Zhiqiang Qin5, Xian Gao2,6, Kai Xu1, Xiaokai Shi1, Shenglin Gao1, Shouyong Liu3, Kai Zhu3, Wei Wang3, Li Zuo1, Lifeng Zhang1, Wei Zhang3. 1. Department of Urology, The Affiliated Changzhou No. 2 People's Hospital of Nanjing Medical University, Changzhou, China. 2. Graduate School of Nanjing Medical University, Nanjing, China. 3. Department of Urology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China. 4. Department of Urology, Taizhou People's Hospital, The Fifth Affiliated Hospital of Medical School of Nantong University, Taizhou, China. 5. Department of Urology and Transplantation, Nanjing First Hospital, Nanjing Medical University, Nanjing, China. 6. Department of Oncology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
Abstract
BACKGROUND: 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: RT-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. RESULTS: miR-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. CONCLUSIONS: In 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.
BACKGROUND: 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: RT-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. RESULTS: miR-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. CONCLUSIONS: In 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.
Authors: G Bergers; R Brekken; G McMahon; T H Vu; T Itoh; K Tamaki; K Tanzawa; P Thorpe; S Itohara; Z Werb; D Hanahan Journal: Nat Cell Biol Date: 2000-10 Impact factor: 28.824
Authors: Brian D Nicholson; Nicholas R Jones; Andrew Protheroe; Johnson Joseph; Nia W Roberts; Ann Van den Bruel; Thomas R Fanshawe Journal: Cancer Epidemiol Date: 2019-01-15 Impact factor: 2.984