Phenotype characterization of human melanoma cells resistant to dabrafenib

Cordaro,Fabiola Gilda; De Presbiteris,Anna Lisa; Camerlingo,Rosa; Mozzillo,Nicola; Pirozzi,Giuseppe; Cavalcanti,Ernesta; Manca,Antonella; Palmieri,Giuseppe; Cossu,Antonio; Ciliberto,Gennaro; Ascierto,Paolo A.; Travali,Salvatore; Patriarca,Eduardo J.; Caputo,Emilia

doi:10.3892/or.2017.5963

Phenotype characterization of human melanoma cells resistant to dabrafenib

Authors:
- Fabiola Gilda Cordaro
- Anna Lisa De Presbiteris
- Rosa Camerlingo
- Nicola Mozzillo
- Giuseppe Pirozzi
- Ernesta Cavalcanti
- Antonella Manca
- Giuseppe Palmieri
- Antonio Cossu
- Gennaro Ciliberto
- Paolo A. Ascierto
- Salvatore Travali
- Eduardo J. Patriarca
- Emilia Caputo
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Affiliations: Institute of Genetics and Biophysics (IGB), A. Buzzati-Traverso, CNR, I-80131 Naples, Italy, Istituto Nazionale Tumori Fondazione G. Pascale, I-80131 Naples, Italy, Unit of Cancer Genetics, Institute of Biomolecular Chemistry, CNR, I-07100 Sassari, Italy, Unit of Pathology, Hospital-University Health Unit (AOU), I-07100 Sassari, Italy, Department of Biomedical and Biotechnological Sciences-BIOMETEC, University of Catania, I-95100 Catania, Italy
Published online on: September 18, 2017 https://doi.org/10.3892/or.2017.5963
Pages: 2741-2751
Copyright: © Cordaro et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

In the present study, the phenotype of melanoma cells resistant to dabrafenib (a B-RAF inhibitor) was investigated, to shed more light on melanoma resistance to B-RAF inhibition. Melanoma cells resistant to dabrafenib were generated using 3 different cell lines, A375, 397 and 624.38, all carrying B-RAFV600E, and they were characterized by cytofluorometric analysis, Ion Torrent technology, immunofluorescence and biochemistry. All dabrafenib-resistant cells showed, in addition to a re-activation of MAPK signaling, morphological changes compared to their sensitive counterparts, accompanied by an increase in CD90 (mesenchymal marker) expression and a decrease in E-cadherin (epithelial marker) expression, suggesting an epithelial-to-mesenchymal-like phenotypic transition. However, melanoma cells with TGF-β1-induced epithelial-to-mesenchymal transition (EMT) were more sensitive to dabrafenib treatment compared to the sensitivity noted in the non-TGF‑β1‑induced EMT melanoma cells, suggesting that TGF-β1-induced EMT was not associated with dabrafenib resistance. Although dabrafenib-resistant cells exhibited increased cell motility and E-cadherin/vimentin reorganization, as expected in EMT, all of them showed unvaried E-cadherin mRNA and unchanged Snail protein levels, while Twist1 protein expression was decreased with the exception of A375 dabrafenib-resistant melanoma cells, where it was unaffected. These findings suggest a distinct active EMT-like process adopted by melanoma cells under drug exposure. Furthermore, dabrafenib-resistant cells exhibited stem cell-like features, with Oct4 translocation from the cytoplasm to peri-nuclear sites and nuclei, and increased CD20 expression. In conclusion, our data, in addition to confirming that resistance to dabrafenib is dependent on re-activation of MAPK signaling, suggest that this resistance is linked to a distinct active EMT-like process as well as stem-cell features adopted by melanoma cells.

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