Paulo Ricardo da Silva Gomes1,2, Patricia Candido1, Vitória Ghazarian1, Juliana A Camargo1, Vanessa R Guimarães1, Guilherme L Gonçalves3, Poliana Romão1, Iran A Silva1, Miguel Srougi1,4, William C Nahas5, Kátia R Leite1, Sabrina T Reis1, Ruan Pimenta1,4, Nayara Izabel Viana6,7. 1. Laboratorio de Investigação Médica 55 (LIM55), Faculdade de Medicina, Hospital das Clinicas HCFMUSP, Universidade de Sao Paulo, São Paulo, SP, Brazil. 2. Faculdade de Medicina, Universidade Federal do Pará, Belém, PA, Brazil. 3. Laboratory of Renal Physiology, Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of Sao Paulo, São Paulo, Brazil. 4. D'Or Institute for Research and Education (IDOR), São Paulo, Brazil. 5. Uro-Oncology Group, Urology Department, University of Sao Paulo Medical School and Institute of Cancer Estate of Sao Paulo (ICESP), São Paulo, Brazil. 6. Laboratorio de Investigação Médica 55 (LIM55), Faculdade de Medicina, Hospital das Clinicas HCFMUSP, Universidade de Sao Paulo, São Paulo, SP, Brazil. niviana@usp.br. 7. Universidade do Estado de Minas Gerais - UEMG, Passos, MG, Brazil. niviana@usp.br.
Abstract
BACKGROUND: Bladder cancer is the leading transitional cell carcinoma affecting men and women with high morbidity and mortality rates, justifying the need to develop new molecular target therapies using microRNAs. This study aimed to evaluate the behavior of the T24 cell line after transfection with miR-Let-7c precursor mimic through invasion, migration, apoptosis, and cell cycle assays. METHODS AND RESULTS: T24 cell was transfected with the Let-7c mimic and its respective control and evaluated after 24 h. The expression levels of miR-Let-7c were analyzed by qPCR. We performed wound healing, Matrigel and flow cytometry, apoptosis, and cell cycle assays to determine its effect on cellular processes. Cells transfected with miR-Let-7c showed increased apoptosis rates (p = 0.019), decreased migration 24 h (p = 0.031) and 48 h (p = 0.0006), invasion potential (p = 0.0007), and cell proliferation (p = 0.002). CONCLUSIONS: Our results demonstrate that miR-Let-7c can act in different pathways of the carcinogenic cellular processes of muscle-invasive urothelial carcinoma cells, inhibiting cell proliferation and increasing apoptosis levels, consequently limiting their invasion potential. However, further studies should be carried out better to elucidate this microRNA's role in high-grade urothelial carcinomas and unveil which targets this microRNA may present, which are intrinsically related to the cancer survival pathways.
BACKGROUND: Bladder cancer is the leading transitional cell carcinoma affecting men and women with high morbidity and mortality rates, justifying the need to develop new molecular target therapies using microRNAs. This study aimed to evaluate the behavior of the T24 cell line after transfection with miR-Let-7c precursor mimic through invasion, migration, apoptosis, and cell cycle assays. METHODS AND RESULTS: T24 cell was transfected with the Let-7c mimic and its respective control and evaluated after 24 h. The expression levels of miR-Let-7c were analyzed by qPCR. We performed wound healing, Matrigel and flow cytometry, apoptosis, and cell cycle assays to determine its effect on cellular processes. Cells transfected with miR-Let-7c showed increased apoptosis rates (p = 0.019), decreased migration 24 h (p = 0.031) and 48 h (p = 0.0006), invasion potential (p = 0.0007), and cell proliferation (p = 0.002). CONCLUSIONS: Our results demonstrate that miR-Let-7c can act in different pathways of the carcinogenic cellular processes of muscle-invasive urothelial carcinoma cells, inhibiting cell proliferation and increasing apoptosis levels, consequently limiting their invasion potential. However, further studies should be carried out better to elucidate this microRNA's role in high-grade urothelial carcinomas and unveil which targets this microRNA may present, which are intrinsically related to the cancer survival pathways.
Authors: Nelson Dip; Sabrina T Reis; Luciana S Timoszczuk; Nayara I Viana; Camila B Piantino; Denis R Morais; Caio M Moura; Daniel K Abe; Iran A Silva; Miguel Srougi; Marcos F Dall'Oglio; Katia R M Leite Journal: J Urol Date: 2012-09-20 Impact factor: 7.450
Authors: Lisa Haddick; Wei Zhang; Sören Reinhard; Karin Möller; Hanna Engelke; Ernst Wagner; Thomas Bein Journal: Pharmaceutics Date: 2020-06-02 Impact factor: 6.321