LncRNA OGFRP1 promotes angiogenesis and epithelial-mesenchymal transition in colorectal cancer cells through miR-423-5p/CTCF axis |
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| Institution: | 1. Research Center of Tropical and Infectious Diseases, Kerman University of Medical Sciences, Kerman, Iran;2. Department of Clinical Biochemistry, Afzalipour Faculty of Medicine, Kerman University of Medical Sciences, Kerman, Iran;3. Neuroscience Research Center, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran;4. Endocrinology and Metabolism Research Center, Institute of Basic and Clinical Physiology Sciences, Kerman University of Medical Sciences, Kerman, Iran;5. Clinical Research Development Center of Imam Khomeini Hospital, Jiroft University of Medical Sciences, Jiroft, Iran;6. Department of Cardiology, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran;7. Department of Medical Immunology, Afzalipour Faculty of Medicine, Kerman University of Medical Sciences, Kerman, Iran;8. Department of Medical Microbiology (Bacteriology & Virology), Afzalipour Faculty of Medicine, Kerman University of Medical Sciences, Kerman, Iran;9. Physiology Research Center, Institute of Basic and Clinical Physiology Sciences, Kerman University of Medical Sciences, Kerman, Iran;1. Department of Respiratory Medicine, The First People’s Hospital of Zunyi (The Third Affiliated Hospital of Zunyi Medical University), 563000 Guizhou, China;2. The Third Hospital of Mianyang, Sichuan Mental Health Center, 621000 Sichuan, China;3. School of Public Health, Zunyi Medical University, Zunyi 563000, Guizhou, China;4. Scientific Research Center, The First People''s Hospital of Zunyi (The Third Affiliated Hospital of Zunyi Medical University), 563000 Guizhou, China;5. Department of Respiratory Geriatrics and Otolaryngology, Chongqing Public Health Medical, Chongqing 400030, China;6. Zunyi Medical University, Zunyi 563000, Guizhou, China;1. Laboratory of Molecular Biology and Gene Expression (LABMEG), Federal University of Alagoas (UFAL), Arapiraca, Brazil;2. Faculty of Medicine, Federal University of Alagoas (UFAL), Arapiraca, Brazil;3. Post-graduation Program in Health Science, Institute of Biological Sciences and Health (ICBS), Federal University of Alagoas (UFAL), Maceió, Brazil;4. Crystallography Laboratory, Federal University of Alagoas (UFAL), Maceió, Brazil;1. Division of Bacteriology, ICMR-National Institute of Cholera and Enteric Diseases, P-33 CIT Road, Scheme-XM, Beliaghata, Kolkata 700010, India;2. Division of Electron Microscopy, ICMR-National Institute of Cholera and Enteric Diseases, P-33 CIT Road, Scheme-XM, Beliaghata, Kolkata 700010, India;1. School of Basic Medical Sciences, Chengdu University of Traditional Chinese Medicine, Chengdu, China;2. Department of Gynecology, West China Second University Hospital, Sichuan University, Chengdu, China;1. Immunol. & Cell Culture Labs, Dept. Glycoconjugate Immunochemistry, Center for Glycomics, Institute of Chemistry, Slovak Academy of Sciences, Bratislava, Slovakia;2. Dept. Clin. Immunol .and Allergy, Oncology Institute of St. Elisabeth and Slovak Medical University, Bratislava, Slovakia |
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| Abstract: | ObjectiveTo investigate the mechanism of lncRNA OGFRP1 affecting angiogenesis and epithelial-mesenchymal transition (EMT) in colorectal cancer (CRC) and provide a new target for the treatment of CRC.MethodsThe expressions of OGFRP1, miR-423-5p, and CTCF were measured in CRC cell lines (HT29, LoVo, HCT116, SW620, and SW480) and normal colonic epithelial cells NCM460. Gain and loss of function experiments were performed on HCT116 and SW620 cells, after which the proliferation, apoptosis, EMT, invasion, and migration of the cells were measured using CCK-8 and colony formation assays, flow cytometry, Western blotting, Transwell, and scratch assay. The transfected cells were incubated with human umbilical vein endothelial cells (HUVECs) to assess angiogenesis using tube formation assay. ELISA was performed to detect VEGF in the conditioned medium of HCT116 and SW620 cells. The interactions among OGFRP1, CTCF and miR-423-5p were validated by dual-luciferase reporter assay.ResultsCRC cell lines had increased expression levels of OGFRP1 and CTCF and a suppressed expression level of miR-423-5p when compared with NCM460 cells. Suppression on OGFRP1 or CTCF and overexpression of miR-423-5p led to inhibited proliferation, EMT, invasion and migration and increased apoptosis of HCT116 and SW620 cells. HUVECs incubated with cells transfected with si-OGFRP1, si-CTCF or miR-423-5p mimic had suppressed angiogenesis ability. The effect of OGFRP1 suppression in CRC cells could be counteracted by miR-423-5p inhibition. Both CTCF and OGFRP1 could bind to miR-423-5p.ConclusionOGFRP1 promotes proliferation, EMT, and angiogenesis in CRC through miR-423-5p/CTCF axis. |
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| Keywords: | LncRNA OGFRP1 Colorectal cancer MiR-423-5p CCCTC-binding factor Angiogenesis Epithelial-mesenchymal transition |
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