Global gene expression profiling of human lung epithelial cells after exposure to nanosilver
Ontology highlight
ABSTRACT: In this study, the effect of AgNPs on the gene expression in the human lung epithelial cell line A549, exposed to 12.1 ug/ml AgNPs (EC20) for 24 and 48 hours was compared with the response to control and silver ion (Ag+) treated cells (1.3 ug/ml) using microarray analysis.
Project description:Silver nanoparticles (AgNPs) have shown great potential as therapeutic agents due to their ability to cause apoptotic cell death in cancer cells. However, little knowledge is available regarding the underlying action mechanisms of AgNPs towards multi-drug resistant cancer cells. Herein, we employed quantitative proteomics to investigate the cytotoxic mechanisms of AgNPs on both cisplatin-sensitive (A549 cells) and -resistant (A549/DDP cells) human lung adenocarcinoma and to explore their potential anticancer abilities. We first performed cytotoxicity tests and found that AgNPs exert similar cytotoxic effects on A549 and A549/DDP cells. At the proteome level, A549 and A549/DDP cells responded to AgNPs distinctively and similarly by causing cell apoptosis via upregulating RNA metabolism, suppressing VEGF siganling pathway, repressing p53-mediated pathways, promoting cell cycle arrest, etc. Additionally, AgNPs remarkably induced ROS generation in A549 and A549/DDP cells. The mitotoxicity results further confirmed the effectiveness of AgNPs in hampering mitochondrial function and respiration in A549 and A549/DDP cells. Overall, our investigations showed that AgNPs could effectively induce cell deaths in human lung adenocarcinoma cells regardless of their sensitivities to cisplatin, suggesting that AgNPs could be potentially used in biomedical aspects as an anticancer agent in alleviating the problem of acquired drug resistance in chemotherapy.
Project description:The mode of action of silver nanoparticles (AgNPs) is suggested to be exerted through both Ag+ and AgNP dependent mechanisms. Ingestion is one of the major NP exposure routes, and potential effects are often studied using Caco-2 cells, a well-established model for the gut epithelium. MCF-7 cells are epithelial breast cancer cells with extensive well-characterized toxicogenomics profiles. In the present study we aimed to gain a deeper understanding of the cellular molecular responses in Caco-2 and MCF-7 cells after AgNP exposure in order to evaluate whether epithelial cells derived from different tissues demonstrated similar responses. These insights could possibly reduce the size of cell panels for NP hazard identification screening purposes. AgNPs of 20, 30, 60, and 110 nm, and AgNO3 were exposed for 6h and 24h. AgNPs were shown to be taken up and dissolve intracellularly. Compared with MCF-7 cells, Caco-2 cells showed a higher sensitivity to AgNPs, slower gene expression kinetics, and absence of NP size-dependent responses. However, on a molecular level, no significant differences were observed between the two cell types. Transcriptomic analysis showed that Ag(NP) exposure caused (oxidative) stress responses, possibly leading to cell death in both cell lines. There was no indication for effects specifically induced by AgNPs. Responses to AgNPs appeared to be induced by silver ions released from the AgNPs. In conclusion, differences in mRNA responses to AgNPs between Caco-2 and MCF-7 cells were mainly related to timing and magnitude, but not to a different underlying mechanism. In total 73 samples are analyzed (24 different samples all n=3; except for 1 sample with n=4). Twelve of the 24 different samples are extracted from Caco-2 cells, 6 samples at t=6h and 6 at t=24h The other 12 different of the 24 samples are extracted from MCF-7 cells, 6 samples at t=6h and 6 at t=24h For each cell type and each timepoint one of the 6 samples was a negative control sample
Project description:To identify genes and pathways involved in AgNPs and Ag ion toxicity, mRNA microarray analysis was conducted on human Jurkat T cells. The results indicate that more DEGs were induced by AgNPs than by Ag ion and AgNPs induced gene expression were not clustered with control and Ag ion induced ones. DEG analysis indicated that metallothionein (MT) 2A, 1H, 1F, and 1A and endonucleases G like 1 (ENDOGL1) were upregulated by AgNPs exposure more than 2 folds compared to control. Jurkat T cells were exposed to 0.2 mg/L of AgNPs and Ag ions for 24 h. After treatment, total RNA was extracted and microarray was conducted on control, AgNPs treated and Ag ion treated Jurktat T cells. Microarray analysis were performed in triplicate. Jurkat T cells were exposed to 0.2 mg/L of AgNPs and Ag ions for 24 h. After treatment, total RNA was extracted and mi RNA microarray was conducted on control, AgNPs treated and Ag ion treated Jurktat T cells.
Project description:Custom D. magna gene expression microarray (Design ID: 023710, Agilent Technologies)were used to characterise gene expression profiles of Daphnia magna neoantes exposed to silver nanoparticles ( AgNPs ) or silver nitrate ( AgNO3 ) for 24 hours.
Project description:Silver nanoparticles (AgNP) have been reported to penetrate the central nervous system and induce neurotoxicity. However, there is paucity in understanding the toxicity of AgNPs and their effect on the blood-brain barrier (BBB) including the underlying molecular mechanism(s) of action. Using an in vitro BBB model and mass spectrometry based proteomics we investigated the alteration in the proteome pathways of the brain endothelial cells and astrocytes exposed to AgNPs for 24 and 48 hours.
Project description:Silver nanoparticles (AgNPs), which have been used extensively in consuming products and eventually released into the natural environment, have aroused concerns recently because of their potentially harmful effects on human beings following various routes of exposure. As the liver is one of the largest accumulation and deposition sites of circulatory AgNPs, it is important to evaluate the hepatotoxicity induced by AgNPs. However, the acting mechanisms of AgNPs-induced hepatotoxicity are still elusive to a great extent. Herein, we investigated the hepatotoxic effects of AgNPs using a comparative proteomics approach. First, we evaluated the cytotoxicity of different-sized AgNPs and found that the cancerous liver cells were generally more sensitive than the normal liver cells. Next, proteomics results suggested that HepG2 and L02 cells showed distinct adaptive responses upon AgNPs exposure. HepG2 cells respond to stresses by adapting energy metabolism, upregulating metallothionein expression and increasing the expression of antioxidants, while L02 cells protect themselves by increasing DNA repair and macro-autophagy. Besides, mitochondrial ROS has been identified as one of the causes of AgNPs-induced hepatotoxicity. Collectively, our results revealed that hepatic cancer cells and normal cells cope with AgNPs in notably different pathways, providing new insights into mechanisms underlying AgNPs-induced hepatotoxicity.
Project description:To investigate the patterns of global gene expression profiles modulated by the different sized AgNPs and to differentiate their modes of toxicity, zebrafish (Danio rerio) will be exposed to the AgNPs (50, and 150nm) and used for microarray analysis by Agilent Zebrafish Oligo Microarray system. 334 genes overlaped between AgNPs and Ag+ treatments in a total of 7,538 differential expressed genes. Immune response, antigen processing and presentation, response to estradiol stimulus and regulation of RNA metabolic process are most significant GO terms enriched in genes up regulated by four treatments. Neuroactive ligand-receptor interaction pathway was enriched among AgNP 50nm and AgNP 150nm activated genes, and specifically induced by AgNP 50nm include cell cycle and Toll-like receptor signaling pathways. The zebrafish larvae (72hpf) was exposed to AgNPs for 96hour. And then after total RNA extration from the each samples, the AgNPs related gene expression profiles identified using Agilent Zebrafish Oligo Microarray. Significant alterations in gene expression were found for all treatments and many of the gene pathways connected.
Project description:Addressing the challenges of melanoma, including its aggressive nature and treatment resistance. This study explores the application of silver nanoparticles (AgNPs) synthesized via conventional and green methods. By examining proteomic changes in A375 melanoma cells, we identify key disruptions in cell viability and gene activity. The DIA MS/SWATH MS analysis generates a set of differentially expressed proteins with respect to AgNPs. This allows us to identify new potential regulatory mechanisms and molecular pathways. AgNP exposure leads to oxidative stress, mitochondrial dysfunction, and activation of apoptosis pathways, revealing the potential of AgNPs in melanoma treatment. This comprehensive analysis highlights the promise of eco-friendly nanoparticle synthesis in developing effective anti-cancer therapies.
Project description:Applications for silver nanomaterials in consumer products are rapidly expanding, creating an urgent need for toxicological examination of the exposure potential and ecological effects of silver nanoparticles (AgNPs). The integration of genomic techniques into environmental toxicology has presented new avenues to develop exposure biomarkers and investigate the mode of toxicity of novel chemicals. In the present study we used a 15k oligonucleotide microarray for Daphnia magna, a freshwater crustacean and common indicator species for toxicity, to differentiate between particle specific and ionic silver toxicity and to develop exposure biomarkers for citrate-coated and PVP-coated AgNPs. Gene expression profiles revealed that AgNO3 and AgNPs have distinct expression profiles suggesting different modes of toxicity. However, the gene expression profiles of the different coated AgNPs were similar revealing similarities in the cellular effects of these two particles. Major biological processes disrupted by the AgNPs include protein metabolism and signal transduction. In contrast, AgNO3 caused a downregulation of developmental processes, particularly in sensory development. Metal responsive and DNA damage repair genes were induced by the PVP AgNPs, but not the other treatments. In addition, two specific biomarkers were developed for the environmental detection of PVP AgNPs; although further verification under different environmental conditions is needed. We exposed Daphnia magna to the 1/10 LC50 and LC25 of citrate coated and PVP-coated Ag nanoparticles and Ag+ as AgNO3 for 24-h. For each exposure condition, we performed 6 replicate exposures with 5 individuals in each. All exposures were compared to a unexposed laboratory control.
Project description:Nanoparticles are compounds of emerging concern with largely unknown risks for human and ecological health. It is crucial to evaluate their potential biological impact to prevent unintended adverse effects on human health and the environment. We analyzed the transcriptional effects of polyvinylpyrrolidone-coated silver nanoparticles (PVP-AgNPs) and silver nitrate (AgNO3) on the fathead minnow (Pimephales promelas) to understand their potential toxicity and adverse outcomes. We also tested the feasibility of the fathead minnow as an alternative species to elucidate potential adverse effects on humans. Fathead minnow females were exposed to either 4 µg/L of AgNO3 or 70 µg/L of PVP-AgNPs for 96h. Microarray analyses were performed on liver and brain. Functional analysis identified potential toxicity pathways and molecular initiating events (MIEs) that were confirmed with functional assays. Data suggested that AgNO3 and PVP-AgNPs had both common and distinct transcriptional effects. The nanoparticles were linked to neurotoxicity and oxidative stress, and identified as a dopamine receptor antagonist. Silver nitrate was also identified as a potential neurotoxicant and was confirmed as adrenergic and cannabinoid receptors antagonist. While silver nitrate and PVP-AgNPs were both potential neurotoxicants, they appeared to act through different MIEs. Fathead minnow is a promising alternative species to elucidate potential adverse effects of relevance to human health. We analyzed the transcriptional effects of polyvinylpyrrolidone-coated silver nanoparticles (PVP-AgNPs) and silver nitrate (AgNO3) on the fathead minnow (Pimephales promelas) to understand their potential toxicity and adverse outcomes. FHM were obtained from Aquatic Biosystems (Fort Collins, CO), held in aerated dechlorinated tap water and fed three times daily with Zeigler® AquaTox Feed Gardners, PA, USA). Fathead minnow females were exposed to either 4 µg/L of AgNO3 or 70 µg/L of PVP-AgNPs (Luna Innovations, Blackburn, VA) for 96h at 24°C ± 1 with a 90% water change at 48 hours. Microarray analyses were performed on liver and brain.