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Toxicology ; 322: 69-77, 2014 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-24857880

RESUMO

We examined the role of miRNAs in DNA damage response in HepG2 cells following exposure to 4-aminobiphenyl (4-ABP). The arylamine 4-ABP is a human carcinogen. Using the Comet assay, we showed that 4-ABP (18.75-300µM) induces DNA damage in HepG2 cells after 24h. DNA damage signaling pathway-based PCR arrays were used to investigate expression changes in genes involved in DNA damage response. Results showed down-regulation of 16 DNA repair-related genes in 4-ABP-treated cells. Among them, the expression of selected six genes (UNG, LIG1, EXO1, XRCC2, PCNA, and FANCG) from different DNA repair pathways was decreased with quantitative real-time PCR (qRT-PCR). In parallel, using the miRNA array, we reported that the expression of 27 miRNAs in 4-ABP-treated cells was at least 3-fold higher than that in the control group. Of these differential 27 miRNAs, the most significant expression of miRNA-513a-5p and miRNA-630 was further validated by qRT-PCR, and was predicted to be implicated in the deregulation of FANCG and RAD18 genes, respectively, via bioinformatic analysis. Both FANCG and RAD18 proteins were found to be down-regulated in 4-ABP-treated cells. In addition, overexpression and knockdown of miRNA-513a-5p and miRNA-630 reduced and increased the expression of FANCG and RAD18 proteins, respectively. Based on the above results, we indicated that miRNA-513a-5p and miRNA-630 could play a role in the suppression of DNA repair genes, and eventually lead to DNA damage.


Assuntos
Compostos de Aminobifenil/toxicidade , Carcinógenos/toxicidade , Reparo do DNA/genética , Expressão Gênica/efeitos dos fármacos , RNA Mensageiro/fisiologia , Western Blotting , Sobrevivência Celular/efeitos dos fármacos , Ensaio Cometa , Meios de Cultura , Dano ao DNA , Humanos , Hibridização In Situ , RNA/biossíntese , RNA/isolamento & purificação , RNA Mensageiro/biossíntese , Reação em Cadeia da Polimerase em Tempo Real
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