Atrazine Affects Phosphoprotein and Protein Expression in MCF-10A Human Breast Epithelial Cells
Atrazine, a member of the 2-chloro-s-triazine family of herbicides, is the most widely used pesticide in the world and often detected in agriculture watersheds. Although it was generally considered as an endocrine disruptor, posing a potential threat to human health, the molecular mechanisms of atra...
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doaj-bd1200c5d30f4b82bdbc9d3cdefbc7c72020-11-24T21:59:10ZengMDPI AGInternational Journal of Molecular Sciences1422-00672014-10-011510178061782610.3390/ijms151017806ijms151017806Atrazine Affects Phosphoprotein and Protein Expression in MCF-10A Human Breast Epithelial CellsPeixin Huang0John Yang1Qisheng Song2Department of Agriculture & Environmental Science, Lincoln University of Missouri, Jefferson City, MO 65120, USADepartment of Agriculture & Environmental Science, Lincoln University of Missouri, Jefferson City, MO 65120, USADivision of Plant Sciences, University of Missouri, Columbia, MO 65211, USAAtrazine, a member of the 2-chloro-s-triazine family of herbicides, is the most widely used pesticide in the world and often detected in agriculture watersheds. Although it was generally considered as an endocrine disruptor, posing a potential threat to human health, the molecular mechanisms of atrazine effects remain unclear. Using two-dimensional gel electrophoresis, we identified a panel of differentially expressed phosphoproteins and total proteins in human breast epithelial MCF-10A cells after being exposed to environmentally relevant concentrations of atrazine. Atrazine treatments for 6 h resulted in differential expression of 4 phosphoproteins and 8 total-proteins as compared to the control cells (>1.5-fold, p < 0.05). MALDI-TOF MS/MS analysis revealed that the differentially expressed proteins belong to various cellular compartments (nucleus, cytosol, membrane) and varied in function, including those regulating the stress response such as peroxiredoxin I, HSP70 and HSP27; structural proteins such as tropomyosin and profilin 1; and oncogenesis proteins such as ANP32A. Six of the 12 identified proteins were verified by quantitative PCR for their transcript levels. The most up-regulated phosphoprotein by atrazine treatment, ANP32A, was further analyzed for its expression, distribution and cellular localization using Western blot and immunocytochemical approaches. The results revealed that ANP32 expression after atrazine treatment increased dose and time dependently and was primarily located in the nucleus. This study may provide new evidence on the potential toxicity of atrazine in human cells.http://www.mdpi.com/1422-0067/15/10/17806AtrazineMCF-10A cells2DEprotein expressionANP32A |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Peixin Huang John Yang Qisheng Song |
spellingShingle |
Peixin Huang John Yang Qisheng Song Atrazine Affects Phosphoprotein and Protein Expression in MCF-10A Human Breast Epithelial Cells International Journal of Molecular Sciences Atrazine MCF-10A cells 2DE protein expression ANP32A |
author_facet |
Peixin Huang John Yang Qisheng Song |
author_sort |
Peixin Huang |
title |
Atrazine Affects Phosphoprotein and Protein Expression in MCF-10A Human Breast Epithelial Cells |
title_short |
Atrazine Affects Phosphoprotein and Protein Expression in MCF-10A Human Breast Epithelial Cells |
title_full |
Atrazine Affects Phosphoprotein and Protein Expression in MCF-10A Human Breast Epithelial Cells |
title_fullStr |
Atrazine Affects Phosphoprotein and Protein Expression in MCF-10A Human Breast Epithelial Cells |
title_full_unstemmed |
Atrazine Affects Phosphoprotein and Protein Expression in MCF-10A Human Breast Epithelial Cells |
title_sort |
atrazine affects phosphoprotein and protein expression in mcf-10a human breast epithelial cells |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1422-0067 |
publishDate |
2014-10-01 |
description |
Atrazine, a member of the 2-chloro-s-triazine family of herbicides, is the most widely used pesticide in the world and often detected in agriculture watersheds. Although it was generally considered as an endocrine disruptor, posing a potential threat to human health, the molecular mechanisms of atrazine effects remain unclear. Using two-dimensional gel electrophoresis, we identified a panel of differentially expressed phosphoproteins and total proteins in human breast epithelial MCF-10A cells after being exposed to environmentally relevant concentrations of atrazine. Atrazine treatments for 6 h resulted in differential expression of 4 phosphoproteins and 8 total-proteins as compared to the control cells (>1.5-fold, p < 0.05). MALDI-TOF MS/MS analysis revealed that the differentially expressed proteins belong to various cellular compartments (nucleus, cytosol, membrane) and varied in function, including those regulating the stress response such as peroxiredoxin I, HSP70 and HSP27; structural proteins such as tropomyosin and profilin 1; and oncogenesis proteins such as ANP32A. Six of the 12 identified proteins were verified by quantitative PCR for their transcript levels. The most up-regulated phosphoprotein by atrazine treatment, ANP32A, was further analyzed for its expression, distribution and cellular localization using Western blot and immunocytochemical approaches. The results revealed that ANP32 expression after atrazine treatment increased dose and time dependently and was primarily located in the nucleus. This study may provide new evidence on the potential toxicity of atrazine in human cells. |
topic |
Atrazine MCF-10A cells 2DE protein expression ANP32A |
url |
http://www.mdpi.com/1422-0067/15/10/17806 |
work_keys_str_mv |
AT peixinhuang atrazineaffectsphosphoproteinandproteinexpressioninmcf10ahumanbreastepithelialcells AT johnyang atrazineaffectsphosphoproteinandproteinexpressioninmcf10ahumanbreastepithelialcells AT qishengsong atrazineaffectsphosphoproteinandproteinexpressioninmcf10ahumanbreastepithelialcells |
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