Some environmental chemicals have been shown to cause liver-toxicity as the result of bioaccumulation. Particularly, fungicides have been shown to cause varying degrees of hepatictoxicity and to disrupt steroid hormone homeostasis in in vivo models. The principal objective of this study was to evalu...
Some environmental chemicals have been shown to cause liver-toxicity as the result of bioaccumulation. Particularly, fungicides have been shown to cause varying degrees of hepatictoxicity and to disrupt steroid hormone homeostasis in in vivo models. The principal objective of this study was to evaluate the liver-toxic responses of environmental chemicals-in this case selected fungicides and parasiticides-in order to determine whether or not this agent differentially affected its toxicogenomic activities in hepatic tumor cell lines. To determine the gene expression profiles of 3 fungicides (triadimefon, myclobutanil, vinclozolin) and 1 parasiticide (dibutyl phthalate), we utilized a modified HazChem human array V2. Additionally, in order to observe the differential alterations in its time-dependent activities, we conducted two time (3 hr, 48 hr) exposures to the respective IC20 values of four chemicals. As a result, we analyzed the expression profiles of a total of 1638 genes, and we identified 70 positive significant genes and 144 negative significant genes using four fungicidic and parasiticidic chemicals, using SAM (Significant Analysis of Microarray) methods (q-value<0.5%). These genes were analyzed and identified as being related to apoptosis, stress responses, germ cell development, cofactor metabolism, and lipid metabolism in GO functions and pathways. Additionally, we found 120 genes among those time-dependently differentially expressed genes, using 1-way ANOVA (P-value<0.05). These genes were related to protein metabolism, stress responses, and positive regulation of apoptosis. These data support the conclusion that the four tested chemicals have common toxicogenomic effects and evidence respectively differential expression profiles according to exposure time.
Some environmental chemicals have been shown to cause liver-toxicity as the result of bioaccumulation. Particularly, fungicides have been shown to cause varying degrees of hepatictoxicity and to disrupt steroid hormone homeostasis in in vivo models. The principal objective of this study was to evaluate the liver-toxic responses of environmental chemicals-in this case selected fungicides and parasiticides-in order to determine whether or not this agent differentially affected its toxicogenomic activities in hepatic tumor cell lines. To determine the gene expression profiles of 3 fungicides (triadimefon, myclobutanil, vinclozolin) and 1 parasiticide (dibutyl phthalate), we utilized a modified HazChem human array V2. Additionally, in order to observe the differential alterations in its time-dependent activities, we conducted two time (3 hr, 48 hr) exposures to the respective IC20 values of four chemicals. As a result, we analyzed the expression profiles of a total of 1638 genes, and we identified 70 positive significant genes and 144 negative significant genes using four fungicidic and parasiticidic chemicals, using SAM (Significant Analysis of Microarray) methods (q-value<0.5%). These genes were analyzed and identified as being related to apoptosis, stress responses, germ cell development, cofactor metabolism, and lipid metabolism in GO functions and pathways. Additionally, we found 120 genes among those time-dependently differentially expressed genes, using 1-way ANOVA (P-value<0.05). These genes were related to protein metabolism, stress responses, and positive regulation of apoptosis. These data support the conclusion that the four tested chemicals have common toxicogenomic effects and evidence respectively differential expression profiles according to exposure time.
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문제 정의
However, some fungicides have been demonstrated to induce varying degrees of hepatic toxicity. The principal objective of this study was to compare the gene expression profiles of four fungicide and parasiticide chemicals, and to identify common expression changes that better characterized toxicities by exposure. The differences in time-dependent responses in human hepatoma cells were also evaluated via this toxicogenomic approach9,10.
제안 방법
We chose to modify and add certain liver-toxicity-related genes, as the four chemicals utilized herein have been shown to evidence varying degrees of hepatic toxicity8. In this study, genomic data from in vitro studies of four fungicidal and parasiticidal chemicals were analyzed, demonstrating that there are regulated hepatic biological functions in active human hepatocytes that are commonly disrupted by these chemicals. These differentially expressed genes and affected functions contributed to our understanding of the liver-toxicity induced by fungicidal and parasiticidal chemicals.
Some fungicides are known to be dangerous to human health, including vinclozolin, which is no longer used3. In this study, we utilized four fungicides and parasiticides-myclobutanil, triadimefon, vinclozolin, and dibutyl phthalate (Figure 1). Myclobutanil and triadimefon are agrichemical fungicides that block ergosterol biosynthesis, and thus control several species and strains of fungi4,5.
In this study, we utilized toxicogenomic analysis, identified common changes in four chemicals, and assessed distinct changes over exposure time. These toxicogenomic approaches were applied to delineate the molecular mode of actions and to assess the risks of environmental toxicants13.
대상 데이터
Dibutyl phthalate (CAS No, 84-74-2), triadimefon (CAS No. 43121-43-3), vinclozolin (CAS No, 50471-44-8), myclobutanil (CAS No. 88671-89-0), Dimethylsulphoxide (DMSO) and 3-(4,5-dimethylthizol-2- yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from the Sigma-Aldrich Chemical Company (USA). RPMI-1640 Culture Medium, Dulbecco’s Phosphate Buffered Saline (PBS) and Fetal Bovine Serum (FBS) were obtained from GIBCOTM (USA).
데이터처리
We selected expressed genes that a FDR-adjusted P value of less than 0.05 according to the ANOVA (test type is Welch’s t-test).
이론/모형
Figure 2. Cell viabilities of 4 chemicals using MTT assay. IC20 values (20% inhibition concentration of control) are respectively 96 μM (triadimefon, box), 72 μM (myclobutanil, circle), 320 μM (vinclozolin, triangle), and 340 μM (dibutyl phthalate (DBP), inverse triangle).
Significant expression changes were analyzed via comparison with the exposed and control groups of all chemicals using the Significant Analysis of Microarray (SAM) method (score>±1, q-value<0.05).
참고문헌 (16)
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