Post-transcriptional regulation of Rad51c by miR-222 contributes cellular transformation

English version

Autoři: Emilio Rojas ^aff001; Monica Martinez-Pacheco ^aff002; Maria Alexandra Rodriguez-Sastre ^aff001; Paulina Ramos-Espinosa ^aff001; Mahara Valverde ^aff001
Působiště autorů: Universidad Nacional Autónoma de México, Instituto de Investigaciones Biomédicas, Departamento de Medicina Genómica y Toxicología Ambiental, Mexico City, C.U., México ^aff001; Center for Genomic Sciences, UNAM, Cuernavaca, Mexico ^aff002
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pone.0221681

Souhrn

DNA repair inhibition has been described as an essential event leading to the initiation of carcinogenesis. In a previous study, we observed that the exposure to metal mixture induces changes in the miR-nome of the cells that was correlated with the sub-expression of mRNA involved in processes and diseases associated with metal exposure. From this analysis, one of the miRNAs that shows changes in its expression is miR-222, which is overexpressed in various cancers associated with exposure to metals. In silico studies showed that a possible target for the microRNA-222 could be Rad 51c, a gene involved in the double-stranded DNA repair. We could appreciate that up-regulation of miR-222 reduces the expression both gene and as a protein expression of Rad51c by RT-PCR and immunoblot, respectively. A luciferase assay was performed to validate Rad51c as miR-222 target. Neutral comet assay was performed in order to evaluate DNA double-strand breaks under experimental conditions. Here, we demonstrate that miR-222 up-regulation, directly regulates Rad51c expression negatively, and impairs homologous recombination of double-strand break DNA repair during the initiation stage of cell transformation. This inhibition triggers morphological transformation in a two-stage Balb/c 3T3 cell assay, suggesting that this small RNA acts as an initiator of the carcinogenesis process.

Klíčová slova:

Gene expression – Transfection – MicroRNAs – Protein expression – DNA damage – DNA repair – Precursor cells – NIH 3T3 cells

Zdroje

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