RESUMEN
The aim of this study was to investigate the effect of hexagonal selenium nanoparticles modified by SiRNA (HSNM-SiRNA) to inhibit epidermal growth factor receptor (EGFR) signaling in Human non-small-cell lung cancer (NSCLC). After synthesis, HSNM-SiRNA and HSNs were separately exposed to NSCLC cell lines (A549, H1299, H520, and H1975), and incubated for 6 h at 37 °C. Next, the expression of NFKB, MYC, STAT, ELK1, and GAPDH was evaluated by western blot and real-time PCR. The percentage of apoptotic cells and cell cycle progression were measured when exposed to HSNM-SiRNA and HSNs. Both western blot and real-time PCR results showed that HSNM-SiRNA could down-regulate the expression of all EGFR signaling genes. The percentage of apoptotic cells was significantly increased in all cell lines when exposed to HSNM-SiRNA (P>0.05). HSNM-SiRNA in A549 and H1299 cells significantly increased the proportion of cells in G1/G0 phase and significantly decreased the proportion of cells in S phase.
Asunto(s)
Receptores ErbB/genética , Receptores ErbB/metabolismo , Nanopartículas del Metal , ARN Interferente Pequeño/genética , Selenio , Transducción de Señal , Apoptosis/genética , Composición de Base , Biomarcadores , Carcinoma de Pulmón de Células no Pequeñas/genética , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Ciclo Celular/genética , Línea Celular Tumoral , Receptores ErbB/química , Regulación Neoplásica de la Expresión Génica , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Nanopartículas del Metal/química , Nanopartículas del Metal/ultraestructura , Modelos Moleculares , Conformación Molecular , ARN Interferente Pequeño/administración & dosificación , ARN Interferente Pequeño/químicaRESUMEN
The aim of this study was to evaluate an engineered nanostructure to silence five important oncogenes, including BAG1, MDM2, Bcl-2, BIRC5 (survivin) and XIAP, in acute myeloid leukemia subtype 2 (AML-M2). The smart nanostructures were functionalized gold nanoparticles (FGNs) containing five antisense oligonucleotides (AOs) and one anti-CD33(+)/CD34(+) aptamer. First, the best AO for each gene was selected with the OligoWalk online software, and then different arrangements of AOs were evaluated with the RNAstructure software. Thereafter, naked gold nanoparticles (NGNs) were synthesized by the reaction of 1000 mm HAuCl4 with 10 µg ml(-1) ascorbic acid. Next, five AOs and one anti-CD33(+)/CD34(+) aptamer were attached to NGNs through serial reactions. Later, 5 ml of heparinized blood samples from five AML-M2 patients were prepared, cancerous cells were isolated and then incubated with three concentrations (75, 150 and 300 µg ml(-1)) each of FGNs, NGNs, gold nanoparticles functionalized with scrambled oligonucleotides (GNFSONs) and doxorubicin. Finally, cell death percentage and gene expressions were measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and real-time PCR, respectively. This study showed that FGNs and doxorubicin led to more cell death compared with NGNs and GNFSONs (P<0.05). Interestingly, all concentrations of FGNs led to a decrease in gene expression. As an important finding, although all concentrations of doxorubicin could also inhibit the expression of genes, FGNs had more effect (P<0.05). Moreover, both NGNs and GNFSONs could silence all genes only at a concentration of 300 µg ml(-1). For BCL2 and XIAP, a dose-dependent pattern was observed, but there was no similar pattern for others.