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Serum Exosomes from Epidermal Growth Factor Receptor (EGFR) Mutated Adenocarcinoma Patients Induce Epithelial to Mesenchymal Transition (EMT)
Description
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A2681 - Serum Exosomes from Epidermal Growth Factor Receptor (EGFR) Mutated Adenocarcinoma Patients Induce Epithelial to Mesenchymal Transition (EMT)
Author Block: P. Nadarajan1, S. Chong2, M. O'Callaghan2, R. Kane3, G. Cooke3, A. Fabre4, M. P. Keane2; 1Respiratory, St Vincent's University Hospital and University College Dublin, Dublin, Ireland, 2Respiratory, St Vincent's University Hospital and University College Dublin, Dublin 4, Ireland, 3University College Dublin, Dublin 4, Ireland, 4Histopathology, St Vincent's University Hospital and University College Dublin, Dublin 4, Ireland.
Serum Exosomes from Epidermal Growth Factor Receptor (EGFR) Mutated Adenocarcinoma Patients Induce Epithelial Mesenchymal Transition (EMT)
RATIONALE
Exosomes are extracellular vesicles with a multi-vesicular endosomal origin that is released by all cell types. They contain protein and genetic information (RNA, DNA and microRNA) and likely reflect the phenotypic state of the cell of origin. They are released in abundance from cancer cells and are involved in cell to cell signalling due to their ability to transport RNA molecules and proteins between cells. Exosomes have been reported to influence epithelial mesenchymal transition (EMT) in pathological states such as metastatic lung cancer. We analysed the effects of exosomes derived from human lung cancer serum on A549 cell line.
METHODS
A549 cells were seeded and cultured in a 12-well plate for 24 hours in 10% foetal bovine serum (FBS) containing growth media. At 24 hours, the cells were washed with PBS and the media was replaced with exosome-depleted serum containing media and cultured for a further 24 hours. Cells were incubated on day 3 with exosomes derived from serum of patients without cancer and patients with adenocarcinoma, both wild-type and EGFR mutated, for a further 48 hours. Cells were scraped from the 12-well plate for preparation of cell lysates on day 5 and examined for markers of EMT such as E-cadherin, ZO-1 and Vimentin using Western blotting.
RESULTS
We show that EMT occurred in cells treated with exosomes derived from EGFR-mutated adenocarcinoma serum as demonstrated by a lower expression of epithelial markers E-cadherin and ZO-1 and a higher expression of the mesenchymal marker, Vimentin compared to control. These effects however, were not seen in cells treated with exosomes derived from serum of patients with wild-type adenocarcinoma and serum of patients with no cancer.
CONCLUSION
Our results would suggest that exosomes derived from EGFR-mutated adenocarcinoma serum could be a potential mediator of EMT in recipient cells and may provide insights into the mechanisms of lung cancer progression and metastasis. However, additional studies are required to precisely delineate these mechanisms.
Author Block: P. Nadarajan1, S. Chong2, M. O'Callaghan2, R. Kane3, G. Cooke3, A. Fabre4, M. P. Keane2; 1Respiratory, St Vincent's University Hospital and University College Dublin, Dublin, Ireland, 2Respiratory, St Vincent's University Hospital and University College Dublin, Dublin 4, Ireland, 3University College Dublin, Dublin 4, Ireland, 4Histopathology, St Vincent's University Hospital and University College Dublin, Dublin 4, Ireland.
Serum Exosomes from Epidermal Growth Factor Receptor (EGFR) Mutated Adenocarcinoma Patients Induce Epithelial Mesenchymal Transition (EMT)
RATIONALE
Exosomes are extracellular vesicles with a multi-vesicular endosomal origin that is released by all cell types. They contain protein and genetic information (RNA, DNA and microRNA) and likely reflect the phenotypic state of the cell of origin. They are released in abundance from cancer cells and are involved in cell to cell signalling due to their ability to transport RNA molecules and proteins between cells. Exosomes have been reported to influence epithelial mesenchymal transition (EMT) in pathological states such as metastatic lung cancer. We analysed the effects of exosomes derived from human lung cancer serum on A549 cell line.
METHODS
A549 cells were seeded and cultured in a 12-well plate for 24 hours in 10% foetal bovine serum (FBS) containing growth media. At 24 hours, the cells were washed with PBS and the media was replaced with exosome-depleted serum containing media and cultured for a further 24 hours. Cells were incubated on day 3 with exosomes derived from serum of patients without cancer and patients with adenocarcinoma, both wild-type and EGFR mutated, for a further 48 hours. Cells were scraped from the 12-well plate for preparation of cell lysates on day 5 and examined for markers of EMT such as E-cadherin, ZO-1 and Vimentin using Western blotting.
RESULTS
We show that EMT occurred in cells treated with exosomes derived from EGFR-mutated adenocarcinoma serum as demonstrated by a lower expression of epithelial markers E-cadherin and ZO-1 and a higher expression of the mesenchymal marker, Vimentin compared to control. These effects however, were not seen in cells treated with exosomes derived from serum of patients with wild-type adenocarcinoma and serum of patients with no cancer.
CONCLUSION
Our results would suggest that exosomes derived from EGFR-mutated adenocarcinoma serum could be a potential mediator of EMT in recipient cells and may provide insights into the mechanisms of lung cancer progression and metastasis. However, additional studies are required to precisely delineate these mechanisms.
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