Yejing Hong1, Zhongming Yuan2, Rongzhong Huang2, Zhiqin Wu3, Yongyong Li4. 1. Department of Gerontology, the Fourth Affiliated Hospital, Zhejiang University School of Medicine, YiWu, China. 2. Department of Gerontology, the Second Affiliated Hospital of Chongqing Medical University, Chongqing, China. 3. Department of Gerontology, the Second Affiliated Hospital of Chongqing Medical University, Chongqing, China. Electronic address: 302955@hospital.cqmu.edu.cn. 4. Department of Gerontology, the Second Affiliated Hospital of Chongqing Medical University, Chongqing, China. Electronic address: liyycqmu@163.com.
Abstract
INTRODUCTION AND OBJECTIVES: Long non-coding RNAs (lncRNAs) have garnered interest because of their roles in cancer progression. We aimed to explore the role of the lncRNA embigin pseudogene 1 (EMBP1)-miR-9-5p axis in renal cell carcinoma (RCC). MATERIALS AND METHODS: Expression profiling of miR-9-5p and EMBP1 were performed in RCC cell lines and tumor samples. To evaluate miR-9-5p and EMBP1's role in proliferation, invasion, migration, and colony formation, we performed in vitro assays along with studies in a xenograft tumor model. In silico binding site analysis using the RNA22 algorithm, RNA-immunoprecipitation (RIP), and luciferase reporter assays were used to validate a direct interaction between EMBP1 and miR-9-5p. Changes in key proteins were also analyzed. RESULTS: miR-9-5p was significantly down-regulated, and EMBP1 was significantly up-regulated, in RCC cell lines and tumor tissue. The clinicopathological characteristics of RCC patients significantly correlated with their expression. Overexpression of miR-9-5p or EMBP1 suppression in RCC cell lines significantly retarded their proliferative, migratory, and invasive behavior, in addition to promoting apoptosis and cell-cycle arrest. EMBP1 directly binds to and negatively regulates miR-9-5p. The EMBP1-miR-9-5p axis dysregulated the expression of the epithelial-to-mesenchymal transition (EMT) markers E-cadherin, claudin, and vimentin, the stemness markers KLF4 and Nanog, and the cell cycle checkpoint gene cyclin E2 (CCNE2) and its downstream mediator E2F1. miR-9-5p overexpression or EMBP1 suppression inhibited xenograft tumor growth in vivo, effects that were abrogated by CCNE2 overexpression. CONCLUSIONS: Our findings suggest an important role of the EMBP1/miR-9-5p axis dysregulation in RCC tumor progression.
INTRODUCTION AND OBJECTIVES: Long non-coding RNAs (lncRNAs) have garnered interest because of their roles in cancer progression. We aimed to explore the role of the lncRNA embigin pseudogene 1 (EMBP1)-miR-9-5p axis in renal cell carcinoma (RCC). MATERIALS AND METHODS: Expression profiling of miR-9-5p and EMBP1 were performed in RCC cell lines and tumor samples. To evaluate miR-9-5p and EMBP1's role in proliferation, invasion, migration, and colony formation, we performed in vitro assays along with studies in a xenograft tumor model. In silico binding site analysis using the RNA22 algorithm, RNA-immunoprecipitation (RIP), and luciferase reporter assays were used to validate a direct interaction between EMBP1 and miR-9-5p. Changes in key proteins were also analyzed. RESULTS:miR-9-5p was significantly down-regulated, and EMBP1 was significantly up-regulated, in RCC cell lines and tumor tissue. The clinicopathological characteristics of RCCpatients significantly correlated with their expression. Overexpression of miR-9-5p or EMBP1 suppression in RCC cell lines significantly retarded their proliferative, migratory, and invasive behavior, in addition to promoting apoptosis and cell-cycle arrest. EMBP1 directly binds to and negatively regulates miR-9-5p. The EMBP1-miR-9-5p axis dysregulated the expression of the epithelial-to-mesenchymal transition (EMT) markers E-cadherin, claudin, and vimentin, the stemness markers KLF4 and Nanog, and the cell cycle checkpoint gene cyclin E2 (CCNE2) and its downstream mediator E2F1. miR-9-5p overexpression or EMBP1 suppression inhibited xenograft tumor growth in vivo, effects that were abrogated by CCNE2 overexpression. CONCLUSIONS: Our findings suggest an important role of the EMBP1/miR-9-5p axis dysregulation in RCC tumor progression.
Authors: Maciej Stasiak; Tomasz Kolenda; Joanna Kozłowska-Masłoń; Joanna Sobocińska; Paulina Poter; Kacper Guglas; Anna Paszkowska; Renata Bliźniak; Anna Teresiak; Urszula Kazimierczak; Katarzyna Lamperska Journal: Life (Basel) Date: 2021-12-07