ABSTRACT: MicroRNA (miRNA) plays an important role in various diseases and in cellular and molecular responses to toxicants. In the present study, we investigated differential expression of miRNAs in response to three triazole fungicides (myclobutanil, propiconazole, and triadimefon). The human hepatoma cell line (HepG2) was treated with the above triazoles for 3h or 48h. miRNA-based microarray experiments were carried out using the Agilent human miRNA v13 array. At early exposure (3h), six miRNAs were differentially expressed andat late exposure (48h), three miRNAs were significantly expressed. Overall, this study provides an array of potential biomarkers for the above triazole fungicides. Furthermore, these miRNAs induced by triazoles could be the foundation for the development of a miRNA-based toxic biomarker library that can predict environmental toxicity.
Toxicology 11/2012; · 3.68 Impact Factor
ABSTRACT: MicroRNA (miRNA) are approximately 22nt RNA molecules with the ability to regulate gene expression by interacting with its
target mRNA. Recent studies demonstrated that miRNA are responsible for determining cell fate and also plays an important
role in cellular response to xenobiotics stress and other toxicological phenomenon. Also a number of reports have strengthened
the correlation between altered miRNA expression and various cancers. In this study miRNA and mRNA expression profiling was
carried out in in-vitro differentiating MCF-7 and HepG2 cell line to understand the effect of nonylphenol (NP), an industrially synthesized environmental
toxicant. Analyzing the mRNA-miRNA interaction, we observed correlation between deregulated miRNAs and its predictive differentially
expressed target mRNA. We also observed differential expression of two widely studied miRNAs, miR-21 and miR-34 in these cell
lines. CTCF is found to be the predictive target of expressed miR-21 in MCF-7. In HepG2 cell line expressed MDM2 is found
to be a predictive target of miR-34b. These results support the possible role of miRNA interaction in the expression of its
target genes and also ability of environmental toxicant to deregulate miRNA expression.
KeywordsNonylphenol–mRNA–miRNA–Environmental toxicant–Expression profile
Molecular and Cellular Toxicology 04/2012; 7(3):259-269. · 0.88 Impact Factor
ABSTRACT: It is important to acknowledge the harmful effects of environmental chemicals in human’s lives. The toxic effects of Diethylstilbestrol
(DES), one of the endocrine-disrupting chemicals (EDCs), have been documented in many studies. As expected, DES affect male
gendal hormone as well as female’s; therefore, epigenetic study should be considered. In this study, microarray technology
was used to study harmful effects on the level of genomics, and here, two types of microarray chips- the Agilent mouse genome
4 × 44 K array for gene expression profiling and the Agilent mouse miRNA v13 for miRNA expression profiling-was used to study
the relation between gene and miRNA expression profiles. As a result, we identified 4 miRNAs (miR 203, 350, 421, and 466i)
that were similarly expressed at 3 hrs and 24 hrs of DES treat times. Twenty one genes matched between predicted target for
4 miRNAs and 118 genes expressed similarly. These genes have functions related to cell differentiation and cell cycle. Therefore,
DES affects cellular function and induces toxicity in TM4 cells. In future studies, it is necessary to find more related functions
and mechanisms of DES in the system.
KeywordsDES (diethylstilbestrol)–miRNA expression–mRNA expression–Environmental toxicity
ABSTRACT: Diethylstilbestrol (DES), a synthetic estrogen, was examined for genotoxicity in mouse testicular Sertoli TM4 cells using
an in vitro micronucleus assay and microarray analysis to clarify the molecular mechanisms underlying the genotoxicity of estrogenic
compounds on the male reproductive system. The micronucleus test showed that DES induced genotoxic effects on TM4 cells with
S9 activation. Gene expression profiles were studied in DES-treated cells and positive controls, which were cyclophosphamide
(CPA)-treated TM4 cells, as compared to the negative controls. In total, 349 and 328 genes were identified as being either
up- or down-regulated, with over 2-fold changes, in DES- and CPA-treated TM4 cells, respectively. Biofunction and canonical
pathways of differentially expressed genes were analyzed using Ingenuity Pathways Analysis, which were mainly categorized
as cellular development and growth/proliferation. In addition, genes related to cell cycle regulation, such as Egr1, Far1, Cd44, Wint16, Sox6, Sox14, Dnmt3a, and Hdac11, were differentially expressed in DES-treated TM4 cells. A gene network analysis was also performed. Comprehensive gene expression
profiling of DES-treated TM4 cells provides valuable information to better understand the genotoxic events of estrogenic chemicals
in testicular Sertoli cells.
KeywordsDiethylstilbestrol (DES)-Genotoxic effect-
In vitro micronucleus test-Gene expression profiling-Mouse Sertoli TM4 cells
BioChip journal 04/2012; 4(1):49-56. · 0.86 Impact Factor
ABSTRACT: Presently, it is important to address the harmful effects of environmental chemicals on living organisms. The toxic effects
of bisphenol A (BPA), one of the endocrine disruption chemicals (EDCs), have been documented in many studies since the 1930s.
However, BPA has continued to be used to make polycarbonate plastic, etc. Therefore, it is necessary to conduct toxicogenomic
studies to assess the harmful effects of BPA. In this study, microarray technology was used to study the harmful effects format
the genomic level. We used two types of microarray chips to study of the relationship between gene and miRNA expression profiles,
the Agilent mouse genome 4×44 K array for gene expression profiling and the Agilent mouse miRNA v13 for miRNA expression profiling.
As a result, we identified 37 miRNAs that were 2-fold up-or down-regulated within 24 hrs than 3 hrs of BPA treat times. The
gene expression patterns related to miRNAs were classified into a total of 4 groups. The first and third groups and the second
and fourth groups had similar functions to each other as determined by related gene expression. In the first and third groups
included overexpressed genes that were related to metabolism. The second and fourth groups included overexpressed genes that
were related to reproduction. miRNA expression levels were exchanged for BPA degradation. So, genes of related metabolism
were overexpressed. This result was occurred the side effect that was down-expression of reproduction. Thus, miRNAs were regulated
by BPA, and gene expression was subsequently regulated by miRNAs.
KeywordsToxicogenomics-BPA (bisphenol A)-EDCs (Endocrine-disrupting chemicals)-miRNA (microRNA)-Microarray
BioChip journal 04/2012; 4(1):75-81. · 0.86 Impact Factor
ABSTRACT: Pesticide is a chemical substance, biological agent, antimicrobial, disinfectant or device used against any pest, and it can
be grouped in several different ways. Atrazine is one of the most commonly used herbicides found in the rural environments,
permethrin and prallethrin are pyrethroid insecticides. In this study, we compared the characters of herbicide and insecticide
by cytotoxicity and microarray experiments. In the cytotoxicity tests, we couldn’t find specific chemical features. However,
in the gene expression analysis, we found that insecticides affects to RNA processing and RNA metabolism, and especially,
in the energy metabolism, such as generation of precursor metabolites and energy, and energy derivation by oxidation of organic
compounds, herbicides and insecticides are differently working. This study provides characteristics of two pesticides by checking
the transcriptional change.
KeywordsLiver toxicity–Atrazine–Permethrin–Prallethrin–Functional analysis
Molecular and Cellular Toxicology 04/2012; 6(4):378-383. · 0.88 Impact Factor
ABSTRACT: The endocrine system is a system of glands that secrete hormones to regulate the body. This system is disrupted by endocrine
disrupting chemicals (EDCs), which are similar to sex hormones. These chemicals function as androgen antagonists or estrogen
agonists. To determine the effect of EDCs on the gene expression profile in human cells, we treated HepG2 cells with 3 chemicals
(bisphenol A, 17β-estradiol, vinclozolin), and analyzed common gene expression using a custom-made HazChem human array V3.
The Haz-Chem human array V3 included a total of 1136 genes, all of which were differentially expressed by exposure to VOCs,
PAHs, POPs, and LTCs. Of these genes, 24 genes were commonly expressed after exposure to all 3 chemicals, based on the SAM
(Significant Analysis of Microarray) method (q-value < 0.5%). These genes were analyzed further and found to be involved in
coagulation, hemostasis, wound healing, angiogenesis, homeostasis, cell redox homeostasis, and cell proliferation based on
GO function and pathway networks.
Molecular and Cellular Toxicology 04/2012; 6(1):57-63. · 0.88 Impact Factor
ABSTRACT: Polycyclic aromatic hydrocarbons (PAHs) are one of the most widespread organic pollutants, and they can cause changes in gene
expression. Biochip based microarray technologies are useful for identifying the toxicity of environmental pollutants. In
this study, we performed a microarray experiment to identify the effects of PAHs in human hepatoma cells using the HazChem
human array V3. The cells were exposed for 48 hours to IC20 values of four PAH chemicals (benzo[a]pyrene, dibenzo[a,h]anthracene, 3-methylcholanthrene, benzo[k]fluoranthene). Thirty
of 1136 genes, which were related to the cell cycle based the ontology analysis, were commonly detected when cells were exposed
to the four chemicals. Further analysis of the significantly expressed genes revealed that they affected the spindle check
point. These results suggest that PAHs affect cell cycle regulation in humans.
BioChip journal 04/2012; 4(1):30-34. · 0.86 Impact Factor
ABSTRACT: It is known that some environmental chemicals affect the human endocrine system. The harmful effects of endocrine disrupting chemical (EDC) nonylphenol (NP) have been studied since the 1980s. It is known that NP adversely affects physiological functions by mimicking the natural hormone 17 beta-estradiol. In the present study, we analyzed the expression of miRNAs and their target genes in mouse Sertoli TM4 cells to better understand the regulatory roles of miRNAs on Sertoli cells after NP exposure.
Mouse TM4 Sertoli cells were treated with NP for 3 or 24 h, and global gene and miRNA expression were analyzed using Agilent mouse whole genome and mouse miRNA v13 arrays.
We identified genes that were > 2-fold differentially expressed in NP-treated cells and control cells (P < 0.05) and analyzed their functions through Gene Ontology analysis. We also identified miRNAs that were differentially expressed in NP-treated and control cells. Of the 186 miRNAs the expression of which differed between NP-treated and control cells, 59 and 147 miRNAs exhibited 1.3-fold increased or decreased expression at 3 and 24 h, respectively. Network analysis of deregulated miRNAs suggested that Ppara may regulate the expression of certain miRNAs, including miR-378, miR-125a-3p miR-20a, miR-203, and miR-101a, after exposure to NP. Additionally, comprehensive analysis of predicted target genes for miRNAs showed that the expression of genes with roles in cell proliferation, the cell cycle, and cell death were regulated by miRNA in NP-treated TM4 cells. Levels of expression of the miRNAs miR-135a* and miR-199a-5p were validated by qRT-PCR. Finally, miR-135a* target gene analysis suggests that the generation of reactive oxygen species (ROS) following exposure to NP exposure may be mediated by miR-135a* through regulation of the Wnt/beta-catenin signaling pathway.
Collectively, these data help to determine NP's actions on mouse TM4 Sertoli cells and increase our understanding of the molecular mechanisms underlying the adverse effects of xenoestrogens on the reproductive system.
Reproductive Biology and Endocrinology 09/2011; 9:126. · 2.05 Impact Factor
ABSTRACT: Thioacetamide (TA) is a potent hepatotoxicant known to affect liver metabolism, inhibit mRNA transport and induce immune suppression. The genetic mechanism underlining this biological toxic compound is well understood using microarray technology. Thus, we used high-throughput rat genome oligonucleotide microarrays containing approximately 22,000 genes to investigate the genetic components of TA-related cytotoxicity in WB-F344 rat liver epithelial (WB-F344) cells. We treated cells with TA (two concentrations over five time periods, ranging from 1 to 24 hr), isolated total RNA at 1, 3, 6, 12 and 24 hr following TA treatment and hybridized the RNA to microarrays. Clustering analysis distinguished two groups of genes, early (1 and 3 hr) and late (6, 12 and 24 hr) phase genes. In total, 2,129 and 2,348 differentially-expressed genes were identified following treatment with low and high concentrations of TA, respectively. A common set of 1,229 genes that were differentially expressed following treatment with both low (1,000 muM) and high (10,000 muM) concentrations of TA had similar expression patterns. Interestingly, 1,410 genes at the low concentration and 1,858 genes at the high concentration were differentially expressed in the early phases, suggesting that these genes associated with the early response to TA may be useful as early markers of hepatotoxicity.
Journal of Veterinary Medical Science 07/2009; 71(6):719-27. · 0.85 Impact Factor
ABSTRACT: The purpose of this study was to apply the multiplex bead array as a diagnostic tool for male infertility. The multiplex bead array offers a new platform in high-throughput nucleic acid detection. Six loci, including sex-determining regions on the Y (SRY) chromosome as a control and five sequence-tagged sites (STS) in azoospermia-factor regions, were used in this system. Extracted genomic DNA from infertile male blood was used for multiplex polymerase chain reaction (PCR). After multiplex PCR using specific Cy3-labeled primer sets, the PCR product was hybridized with capture probes. A multiplexed PCR-liquid bead was arrayed for simultaneous detection using the Luminex system. This assay system correctly identified the presence or deletion of the Y chromosome. Therefore, this method provides a sensitive and high-throughput method for probing the deletion of the Y chromosome, and offers a completely new approach to male infertility screening.
Molecular and Cellular Probes 05/2008; 22(2):76-82. · 2.08 Impact Factor
ABSTRACT: The recent DNA microarray technology enables us to understand a large number of gene expression profiling. The technology has potential possibility to comprehend mechanism of multiple genes were related to compounds which have toxicity in biological system. So, the toxicogenomics through this technology may be very powerful for understanding the effect of unknown toxic mechanisms in biological system. We have studied that the effect of compounds related to hepatotoxin in vivo system using DNA microarray and classified chemicals which have been well characterized. We have studied three compounds; 2 peroxisome proliferators: Clofibrate (ethyl-p-chlorophenoxyisobutyrate), gemfibrozil (5-2[2,5-dimethyl-phenoxy]2-2-dimethyl-pentanonic), and an antiepileptic drug: phenytoin (5,5-diphenylhydantoin). Male Sprague-Dawely VAF(+) albino rats of 5-6 weeks old were treated with each compound for 24 hr and 2 weeks. 4.8 K cDNA microarray in house has been used for gene expression profiling. We found that the clustering of gene expression had similarity like as the toxic phenotype of compounds.
Journal of Veterinary Medical Science 12/2004; 66(11):1329-33. · 0.85 Impact Factor