Abstract
Exposure to particulate matter (PM) has been related to the onset of adverse health effects including lung cancer, but the underlying molecular mechanisms are still under investigation. Epithelial-to-mesenchymal transition (EMT) is regarded as a crucial step in cancer progression. In a previous study, we reported EMT-related responses in the human bronchial epithelial cell line HBEC3-KT, exposed to Milan airborne winter PM2.5. We also found a strong modulation of SERPINB2, encoding for the PAI-2 protein and previously suggested to play an important role in cancer. Here we investigate the role of SERPINB2/PAI-2 in the regulation of EMT-related effects induced by PM exposure in HBEC3-KT. PM exposure (up to 10 µg/cm2) increased SERPINB2 expression, reduced cell migration and induced morphological alterations in HBEC3-KT. Changes in actin structure and cadherin-1 relocalization were observed in PM-exposed samples. Knockdown of SERPINB2 by siRNA down-regulated the CDH1 gene expression, as well as PAI-2 and cadherin-1 protein expression. SERPINB2 knockdown also increased cell migration rate, and counteracted the PM-induced reduction of cell migration and alteration of cell morphology. SERPINB2 was found to be greatly down-regulated in a HBEC2-KT transformed cell line, supporting the importance of this gene in the regulation of EMT. In conclusion, here we show that PAI-2 regulates CDH1 gene/cadherin-1 protein expression in bronchial HBEC3-KT cells, and this mechanism might be involved in the regulation of cell migration. SERPINB2 down-regulation should be considered part of EMT, and the over-expression of SERPINB2 in PM-exposed samples might be interpreted as an initial protective mechanism.
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Acknowledgements
The authors want to thank Øivind Skare for the support with the statistical analyses of the migration data.
Funding
This study has been supported by the grants of the Italian Ministry of Foreign Affairs and International Cooperation (MAECI) to PM (Proj. ID PGR00786).
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EL performed the experiments and wrote the manuscript. EL and SM discussed and interpreted the findings. EL, SM and PM designed the project. SM, PM and MC supervised the project and revised the manuscript. AB established the transformed cell line T2KT-CSC-H. All authors have read and approved the final manuscript.
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Longhin, E., Camatini, M., Bersaas, A. et al. The role of SerpinB2 in human bronchial epithelial cells responses to particulate matter exposure. Arch Toxicol 92, 2923–2933 (2018). https://doi.org/10.1007/s00204-018-2259-z
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DOI: https://doi.org/10.1007/s00204-018-2259-z