Gabriela Díaz-Piña1, Rosa Ma Ordoñez-Razo2, Eduardo Montes3, Ignacio Páramo3, Carina Becerril1, Alfonso Salgado1, J Alfredo Santibañez-Salgado4, Mariel Maldonado1, Victor Ruiz5,6. 1. Departamento de Investigación en Fibrosis Pulmonar, Instituto Nacional de Enfermedades Respiratorias "Ismael Cosío Villegas", Calz. Tlalpan 4502, Col. Sección XVI, 14080, Mexico City, Mexico. 2. Hospital de Pediatría, Centro Médico Siglo XXI, Instituto Mexicano del Seguro Social, Unidad de Investigación Médica en Genética Humana, Av. Cuauhtémoc 330, Col. Doctores, 06720, Mexico City, Mexico. 3. Clínica de Asma, Instituto Nacional de Enfermedades Respiratorias "Ismael Cosío Villegas", Calz. Tlalpan 4502, Col. Sección XVI, 14080, Mexico City, Mexico. 4. Departamento de Cirugía Experimental, Instituto Nacional de Enfermedades Respiratorias "Ismael Cosío Villegas", Calz. Tlalpan 4502, Col. Sección XVI, 14080, Mexico City, Mexico. 5. Departamento de Investigación en Fibrosis Pulmonar, Instituto Nacional de Enfermedades Respiratorias "Ismael Cosío Villegas", Calz. Tlalpan 4502, Col. Sección XVI, 14080, Mexico City, Mexico. vicoruz@yahoo.com.mx. 6. Laboratorio de Biología Molecular, Instituto Nacional de Enfermedades Respiratorias "Ismael Cosío Villegas", Calz. Tlalpan 4502, Col. Sección XVI, 14080, Mexico City, Mexico. vicoruz@yahoo.com.mx.
Abstract
INTRODUCTION: microRNAs (miRNAs) are small non-coding 1RNAs that post-transcriptionally regulate gene expression. Recent evidence shows that adenosine deaminases that act on RNA (ADAR) can edit miRNAs. miRNAs are involved in the development of different diseases, such as idiopathic pulmonary fibrosis (IPF). In IPF, about 40% of the miRNAs are differentially expressed with respect to controls. Among these miRNAs, miRNA-21 has been found over-expressed in IPF and its targets are anti-fibrosing molecules such as PELI1 and SPRY2. The objective of this study is to determine the role of ADAR1 and 2 on the expression of miRNA-21 in human lung fibroblasts trough quantification of gene expression, protein levels, and overexpression of ADAR1 and 2. METHODS: Six control and six fibrotic primary fibroblast cell cultures were used for RNA extraction, ADAR1, ADAR2, PELI1, SPRY2, miRNA-21, and pri-miRNA-21 expression was measured. Subsequently, two fibrotic fibroblast cultures were used for overexpression of ADAR1 and ADAR2, and they were stimulated with TGFβ1. Real-time PCR and Western blot were performed. RESULTS: ADAR1 is significantly downregulated in IPF fibroblasts; the overexpression of ADAR1 and ADAR2 reestablishes the expression levels of miRNA-21, PELI1, and SPRY2 in fibroblasts of patients with IPF. CONCLUSION: These changes in the processing of miRNAs have great value in pathology diagnosis, including lung diseases, and play an important role in the understanding of molecular mechanisms involved in the development of different pathologies, as well as representing new therapeutic targets.
INTRODUCTION: microRNAs (miRNAs) are small non-coding 1RNAs that post-transcriptionally regulate gene expression. Recent evidence shows that adenosine deaminases that act on RNA (ADAR) can edit miRNAs. miRNAs are involved in the development of different diseases, such as idiopathic pulmonary fibrosis (IPF). In IPF, about 40% of the miRNAs are differentially expressed with respect to controls. Among these miRNAs, miRNA-21 has been found over-expressed in IPF and its targets are anti-fibrosing molecules such as PELI1 and SPRY2. The objective of this study is to determine the role of ADAR1 and 2 on the expression of miRNA-21 in human lung fibroblasts trough quantification of gene expression, protein levels, and overexpression of ADAR1 and 2. METHODS: Six control and six fibrotic primary fibroblast cell cultures were used for RNA extraction, ADAR1, ADAR2, PELI1, SPRY2, miRNA-21, and pri-miRNA-21 expression was measured. Subsequently, two fibrotic fibroblast cultures were used for overexpression of ADAR1 and ADAR2, and they were stimulated with TGFβ1. Real-time PCR and Western blot were performed. RESULTS:ADAR1 is significantly downregulated in IPF fibroblasts; the overexpression of ADAR1 and ADAR2 reestablishes the expression levels of miRNA-21, PELI1, and SPRY2 in fibroblasts of patients with IPF. CONCLUSION: These changes in the processing of miRNAs have great value in pathology diagnosis, including lung diseases, and play an important role in the understanding of molecular mechanisms involved in the development of different pathologies, as well as representing new therapeutic targets.
Entities:
Keywords:
ADAR edition; IPF; miRNA processing; miRNA-21
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