Florin Zaharie1, Mihai Stefan Muresan2, Bobe Petrushev3, Cristian Berce4, Grigore-Aristide Gafencu5, Sonia Selicean5, Ancuta Jurj5, Roxana Cojocneanu-Petric5, Cosmin-Ioan Lisencu6, Laura-Ancuta Pop5, Valentina Pileczki7, Dan Eniu2, Mihai-Andrei Muresan2, Roxana Zaharie8, Ioana Berindan-Neagoe9, Ciprian Tomuleasa10, Alexandru Irimie11. 1. Iuliu Hatieganu University of Medicine and Pharmacy;Dept. of Surgery, Octavian Fodor Regional Institute of Gastroenterology and Hepatology, Cluj-Napoca, Romania. 2. Iuliu Hatieganu University of Medicine and Pharmacy;Dept. of Surgical and Gynecological Oncology, Ion Chiricuta Oncology Institute, Cluj-Napoca, Romania. 3. Dept of Pathology, Emergency County Hospital, Cluj-Napoca, Romania. 4. Animal Facility, Iuliu Hatieganu University of Medicine and Pharmacy, Cluj-Napoca, Romania. 5. Research Center for Functional Genomics and Translational Medicine Iuliu Hatieganu University of Medicine and Pharmacy, Cluj-Napoca, Romania. 6. Iuliu Hatieganu University of Medicine and Pharmacy; Dept. of Surgical and Gynecological Oncology, Ion Chiricuta Oncology Institute, Cluj-Napoca, Romania. 7. Iuliu Hatieganu University of Medicine and Pharmacy;Research Center for Functional Genomics and Translational Medicine Iuliu Hatieganu University of Medicine and Pharmacy, Cluj-Napoca, Romania. 8. Iuliu Hatieganu University of Medicine and Pharmacy;Dept. of Gastroenterology Octavian Fodor Regional Institute of Gastroenterology and Hepatology, Cluj Napoca; Romania. 9. Research Center for Functional Genomics and Translational Medicine Iuliu Hatieganu University of Medicine and Pharmacy, Cluj-Napoca, Romania;Dept. of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. 10. Research Center for Functional Genomics and Translational Medicine Iuliu Hatieganu University of Medicine and Pharmacy;Dept. of Hematology, Ion Chiricuta Oncology Institute, Cluj Napoca, Romania. ciprian.tomuleasa@umfcluj.ro. 11. Iuliu Hatieganu University of Medicine and Pharmacy, Cluj-Napoca, Romania.
Abstract
BACKGROUND AND AIMS: Worldwide, colorectal cancer (CRC) is the third most common cancer in men and second in women. The aim of the current study was to identify whether the miR-375 is indeed down-regulated in metastatic CRC and if it could be considered as a potential minimally invasive prognostic biomarker for CRC. METHODS: Exosomes were isolated and characterized from patients with liver metastasis from CCR. The characterization of exosome was performed using TEM/SEM. HCT116 cells were treated with miR-375 mimic, NSM and miR-375 inhibitor. Functional assays included cell counting assay for 14 days, Matrigel invasion assay, apoptosis assay by flow cytometry using Annexin V-FITC, RT-PCR and Western blotting. RESULTS: Increased proliferation potential was proven for the cells transfected with miR-375 inhibitor, while the miR-375 mimic decreased the cell number. The cells transfected with the miR-375 inhibitor are aggressive and cross the membrane; 3.84% of the cells transfected with the miR-375 inhibitor entered apoptosis, while 6.45% of those transfected with the non-specific mimic were in programmed cell death, less than those transfected with the microRNA. RT-PCR for Bcl-2 expression showed that Bcl-2 is down-regulated for miR-375 inhibitor and up-regulated for the miR-375 mimic, a result confirmed by Western blotting. CONCLUSION: The present study brings to the forefront new data that suggest miR-375 as a new player in controlling the pathways responsible for inhibiting the natural history of CRC tumor cells, via the Bcl-2 pathway.
BACKGROUND AND AIMS: Worldwide, colorectal cancer (CRC) is the third most common cancer in men and second in women. The aim of the current study was to identify whether the miR-375 is indeed down-regulated in metastatic CRC and if it could be considered as a potential minimally invasive prognostic biomarker for CRC. METHODS: Exosomes were isolated and characterized from patients with liver metastasis from CCR. The characterization of exosome was performed using TEM/SEM. HCT116 cells were treated with miR-375 mimic, NSM and miR-375 inhibitor. Functional assays included cell counting assay for 14 days, Matrigel invasion assay, apoptosis assay by flow cytometry using Annexin V-FITC, RT-PCR and Western blotting. RESULTS: Increased proliferation potential was proven for the cells transfected with miR-375 inhibitor, while the miR-375 mimic decreased the cell number. The cells transfected with the miR-375 inhibitor are aggressive and cross the membrane; 3.84% of the cells transfected with the miR-375 inhibitor entered apoptosis, while 6.45% of those transfected with the non-specific mimic were in programmed cell death, less than those transfected with the microRNA. RT-PCR for Bcl-2 expression showed that Bcl-2 is down-regulated for miR-375 inhibitor and up-regulated for the miR-375 mimic, a result confirmed by Western blotting. CONCLUSION: The present study brings to the forefront new data that suggest miR-375 as a new player in controlling the pathways responsible for inhibiting the natural history of CRC tumor cells, via the Bcl-2 pathway.
Authors: Marcelo A Mori; Raissa G Ludwig; Ruben Garcia-Martin; Bruna B Brandão; C Ronald Kahn Journal: Cell Metab Date: 2019-08-22 Impact factor: 27.287
Authors: Erika Larrea; Carla Sole; Lorea Manterola; Ibai Goicoechea; María Armesto; María Arestin; María M Caffarel; Angela M Araujo; María Araiz; Marta Fernandez-Mercado; Charles H Lawrie Journal: Int J Mol Sci Date: 2016-04-27 Impact factor: 5.923