Research Papers: Pathology:
Donor-derived stem-cells and epithelial mesenchymal transition in squamous cell carcinoma in transplant recipients
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Abstract
Laurence Verneuil1,2,3,4, Christophe Leboeuf1,2, Guilhem Bousquet1,2, Charlotte Brugiere1,2,3,4, Morad Elbouchtaoui1,2,5, Louis-François Plassa2, Marie-Noelle Peraldi2,6, Celeste Lebbé2,7, Philippe Ratajczak1,2 and Anne Janin1,2,5
1 INSERM, UMR_S1165, Paris, F-75010, France
2 Department of Pathology, Université Paris Diderot, UMR_S1165, F-75010 Paris, France
3 Department of Dermatology, CHU Caen, Caen, F-14033, France
4 Université de Caen Normandie, Medical School, Caen, F-14000, France
5 Department of Pathology, AP-HP, Hôpital Saint-Louis, Paris, F-75010, France
6 Department of Nephrology, AP-HP, Hôpital Saint-Louis, Paris, F-75010, France
7 Department of Dermatology, AP-HP, Hôpital Saint-Louis, Paris, F-75010, France
Correspondence to:
Anne Janin, email:
Laurence Verneuil, email:
Keywords: squamous cell carcinoma, stem-cell, epithelial mesenchymal transition, kidney transplant, chimerism, Pathology Section
Received: May 17, 2015 Accepted: November 16, 2015 Published: November 22, 2015
Abstract
Background
Skin squamous-cell-carcinoma (SCC), is the main complication in long-term kidney-transplant recipients, and it can include donor-derived cells. Preclinical models demonstrated the involvement of epithelial mesenchymal transition (EMT) in the progression of skin SCC, and the role of Snail, an EMT transcription factor, in cancer stem-cell survival and expansion.
Here, we studied stem-cells and EMT expression in SCCs and concomitant actinic keratoses (AK) in kidney-transplant recipients.
Methods
In SCC and AK in 3 female recipients of male kidney-transplants, donor-derived Y chromosome in epidermal stem cells was assessed using combined XY-FISH/CD133 immunostaining, and digital-droplet-PCR on laser-microdissected CD133 expressing epidermal cells.
For EMT study, double immunostainings of CD133 with vimentin or snail and slug, electron microscopy and immunostainings of keratinocytes junctions were performed. Digital droplet PCR was used to check CDH1 (E-cadherin) expression level in laser-microdissected cells co-expressing CD133 and vimentin or snail and slug.
The numbers of Y-chromosome were assessed using digital droplet PCR in laser-microdissected cells co-expressing CD133 and vimentin, or snail and slug, and in CD133 positive cells not expressing any EMT maker.
Results
We identified donor-derived stem-cells in basal layers and invasive areas in all skin SCCs and in concomitant AKs, but not in surrounding normal skin.
The donor-derived stem-cells expressed the EMT markers, vimentin, snail and slug in SCCs but not in AKs. The expression of the EMT transcription factor, SNAI1, was higher in stem-cells when they expressed vimentin. They were located in invasive areas of SCCs. In these areas, the expressions of claudin-1 and desmoglein 1 were reduced or absent, and within the basal layer there were features of basal membrane disappearance.
Donor-derived stem cells were in larger numbers in stem cells co-expressing vimentin or snail and slug than in stem cells not expressing any EMT marker.
Conclusion
We identified here donor-derived stem cells within skin SCC in kidney-transplant recipients. They were located in invasive areas of SCC and had EMT characteristics.
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