RESUMO
Histone deacetylases (HDACs) inhibitor is a promising new approach to the treatment of lung cancer therapy via inhibiting cell growth and inducing apoptosis. miR-15a and miR-16-1 are important tumor suppressors through modulating B cell lymphoma 2 (Bcl-2), Cyclin D1, D2, and others. However, whether HDACs inhibitor modulates the expression of miR-15a/16-1 in lung cancer is still unknown. The purpose of our study was to identify a new miRNA-mediated mechanism which plays an important role in the anti-cancer effects of HDACs inhibitor. We found HDACs inhibitors trichostatin A (TSA) and sodium butyrate upregulated the expression of miR-15a/16-1, residing in the host tumor suppressor Dleu2 gene, through increasing the histone acetylation in the region of Dleu2/miR-15a/16-1 promoter in lung cancer cells. Moreover, among class Ι HDACs subtypes, only knockdown of HDAC3 by specific siRNA increased the hyperacetylation of Dleu2/miR-15a/16-1 promoter region and finally resulted in the upregulation of miR-15a/16-1. Furthermore, overexpression of miR-15a/16-1, which were always deleted or downregulated in lung cancer cells, effectively suppressed cell growth and reduced colony formation. Finally, TSA reduced the expression of Bcl-2, an important survival protein in lung cancer cells, partly through upregulation of miR-15a/16-1. Therefore, this offers a therapeutic strategy that lung cancer patients who exhibit low level of miR-15a/16-1 or high activity of HDACs may benefit from HDACs inhibitor-based therapy.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/genética , Inibidores de Histona Desacetilases/farmacologia , Histona Desacetilases/metabolismo , Ácidos Hidroxâmicos/farmacologia , MicroRNAs/genética , Proteínas Supressoras de Tumor/genética , Acetilação/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Ácido Butírico/farmacologia , Carcinoma Pulmonar de Células não Pequenas/enzimologia , Carcinoma Pulmonar de Células não Pequenas/patologia , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Técnicas de Silenciamento de Genes , Histonas/metabolismo , Humanos , Neoplasias Pulmonares/enzimologia , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , MicroRNAs/metabolismo , Regiões Promotoras Genéticas/genética , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , RNA Longo não Codificante , Transferases , Ensaio Tumoral de Célula-Tronco , Proteínas Supressoras de Tumor/metabolismo , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/genéticaRESUMO
Under the condition of sodium citrate as stabilizer, the gold-selenium (AuSe) nano-alloy was prepared by sodium borohydride reduction procedure, and was modified by single-strand aptamer to obtain an aptamer nano-alloy probe (apta-AuSe) for Hg(II). In pH 6.8 Na2HPO4-NaH2PO4 buffer solution and in the presence of NaCl of 33 mmol L(-1), the Apta-AuSe probe is not aggregation. The apta-AuSe interacts with Hg2+ to form stable double-strand T-Hg(II)-T mismatches and to release AuSe nano-alloy particles from the probe. The released AuSe nano-alloy particles (20:1) aggregated to form bigger clusters that resulted in the resonance scattering (RS) intensity (I590 nm) increasing at 590 nm. The increased intensity delta I590 nm was proportional to the Hg2+ concentration from 1.3 to 1466 nmol L(-1), with a detection limit of 0.74 nmol L(-1). The regress equation was delta I590 nm = 0.603c + 2.0. Thus, a new resonance scattering (RS) spectroscopy of apta-AuSe was applied to the analysis of trace mercury ion. This simple, rapid, selective and sensitive aptamer AuSe nano-alloy RS assay was applied to the determination of Hg2+ in wastewater, with satisfactory results.
