Exposures to environmental chemicals that mimic endogenous hormones are proposed for a number of adverse health effects, including infertility, abnormal prenatal and childhood development and above all cancers. In addition, recently miRNA (micro RNA) has been recognized to play an important role in ...
Exposures to environmental chemicals that mimic endogenous hormones are proposed for a number of adverse health effects, including infertility, abnormal prenatal and childhood development and above all cancers. In addition, recently miRNA (micro RNA) has been recognized to play an important role in various diseases and in cellular and molecular responses to toxicants. In this study, endocrine disrupting environmental toxicant, nonylphenol (NP) was treated to MCF-7 (Human breast cancer cell) and HepG2 (Human hepatocellular liver carcinoma) cell line at 3 hrs and 48 hrs time point and miRNA analysis using $mirVana^{TM}$ miRNA bioarray was performed and compared with total mRNA microarray data for the same cell line and treatment. Robust data quality was achieved through the use of dye-swap. Analysis of microarray data identifies a total of 20 and 11 miRNA expressions at 3 hrs and 48 hrs exposure to NP in MCF-7 cell line and a total of 14 and 47 miRNA expression at 3 hrs and 48 hrs exposure respectively to NP in HepG2 cell line. Expression profiling of the selected miRNA (let-7c, miR-16, miR-195, miR-200b, miR200c, miR-205, and miR-589) reveals changes in the expression of target genes related to metabolism, immune response, apoptosis, and cell differentiation. The present study can be informative and helpful to understand the role of miRNA in molecular mechanism of chemical toxicity and their influence on hormone dependent disease. Also this study may prove to be a valuable tool for screening potential estrogen mimicking pollutants in the environment.
Exposures to environmental chemicals that mimic endogenous hormones are proposed for a number of adverse health effects, including infertility, abnormal prenatal and childhood development and above all cancers. In addition, recently miRNA (micro RNA) has been recognized to play an important role in various diseases and in cellular and molecular responses to toxicants. In this study, endocrine disrupting environmental toxicant, nonylphenol (NP) was treated to MCF-7 (Human breast cancer cell) and HepG2 (Human hepatocellular liver carcinoma) cell line at 3 hrs and 48 hrs time point and miRNA analysis using $mirVana^{TM}$ miRNA bioarray was performed and compared with total mRNA microarray data for the same cell line and treatment. Robust data quality was achieved through the use of dye-swap. Analysis of microarray data identifies a total of 20 and 11 miRNA expressions at 3 hrs and 48 hrs exposure to NP in MCF-7 cell line and a total of 14 and 47 miRNA expression at 3 hrs and 48 hrs exposure respectively to NP in HepG2 cell line. Expression profiling of the selected miRNA (let-7c, miR-16, miR-195, miR-200b, miR200c, miR-205, and miR-589) reveals changes in the expression of target genes related to metabolism, immune response, apoptosis, and cell differentiation. The present study can be informative and helpful to understand the role of miRNA in molecular mechanism of chemical toxicity and their influence on hormone dependent disease. Also this study may prove to be a valuable tool for screening potential estrogen mimicking pollutants in the environment.
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문제 정의
In this study we provide information of how human cell line responds to environmental EDCs. Human breast cancer cell line MCF-7 and Human hepatocellular liver carcinoma cell line HepG2 was treated with nonylphenol at different time point and mRNA expression profiling was carried out using the Nimblegen 4 plex human whole genome array, which contains approximately 24, 000 human genes and miRNA expression is analyzed by mfrVana™ miRNA bioarray.
miRNA expression can be controlled by various chemotherapeutics and chemicals such as ethanol in rodents and humans"시I Xenobiotic-mediated miRNA expression has been directly linked with downstream protein expression and cell proliferation in mice16. These studies demonstrate the potential regulatory functions of miRNAs in different cells and tissues.
제안 방법
A functional analysis of genes showing meaningful expression patterns in each experiment was also conducted. Genes that evidenced expressional changes of over 2 fold as compared to the comparison groups in the functional analysis were analyzed using a gene ontology database.
This normalization method aims to make the distribution of intensities for each array in a set of arrays the same. Data of miRNA expression were analyzed using Gene-Spring GX 7.3.1 (Agilent temologies, CA) and background-adjusted, normalized, and log transformed intensity values were analyzed. Fold change filters included the requirement that the genes be present in at least 200% of controls for p・regulated genes and lower than 50% of controls for down-regulated genes.
대상 데이터
Nonylphenol (NP) was purchased from Sigma-Aldrich in liquid form and stock solution is prepared with DMSO (cat.no.D-2438, 50 mL) purchased from Sigma-Aldrich.
이론/모형
Figure 1. Cell viability after the exposure with nonylphenol was analyzed by the MTT assay. The change in cell death is shown as the % of control.
, WI). Relative signal intensities for each gene were generated using the Robust Multi-Array Average algorithm. The data were processed based on quantile normalization method using the NimbleScan Version 2.
Relative signal intensities for each gene were generated using the Robust Multi-Array Average algorithm. The data were processed based on quantile normalization method using the NimbleScan Version 2.4 (Rmhe NimbleGen, Inc., WI). This normalization method aims to make the distribution of intensities for each array in a set of arrays the same.
성능/효과
2 fold gene expression changed were analyzed by comparing treated with control group in both time point. A total of 389 and 857 genes were differentially expressed in MCF-7 cell line at 3 hrs and 48 hrs exposure to NP respectively compared to 1, 358 and 4, 313 genes expressed in HepG2 cell line at 3 hrs and 48 hrs exposure to NP respectively. Table 1 shows the number of gene upregulated and down regulated in this two cell line.
후속연구
In conclusion, this report will give a new insight into the mechanism by which cells alter their translational profiles in response to toxicity and will highlight fundamental roles of miRNA species in the control of such altered translational events in toxicology.
참고문헌 (22)
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