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


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
doi: 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:

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


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