Long-Term Vemurafenib Exposure Induced Alterations of Cell Phenotypes in Melanoma: Increased Cell Migration and Its Association with EGFR Expression
Acquired resistance during BRAF inhibitor therapy remains a major challenge for melanoma treatment. Accordingly, we evaluated the phenotypical and molecular changes of isogeneic human V600E BRAF-mutant melanoma cell line pairs pre- and post-treatment with vemurafenib. Three treatment naïve...
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MDPI AG
2019-09-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/20/18/4484 |
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| author | Eszter Molnár Tamás Garay Marco Donia Marcell Baranyi Dominika Rittler Walter Berger József Tímár Michael Grusch Balázs Hegedűs |
| author_facet | Eszter Molnár Tamás Garay Marco Donia Marcell Baranyi Dominika Rittler Walter Berger József Tímár Michael Grusch Balázs Hegedűs |
| author_sort | Eszter Molnár |
| collection | DOAJ |
| description | Acquired resistance during BRAF inhibitor therapy remains a major challenge for melanoma treatment. Accordingly, we evaluated the phenotypical and molecular changes of isogeneic human V600E BRAF-mutant melanoma cell line pairs pre- and post-treatment with vemurafenib. Three treatment naïve lines were subjected to in vitro long-term vemurafenib treatment while three pairs were pre- and post-treatment patient-derived lines. Molecular and phenotypical changes were assessed by Sulforhodamine-B (SRB) assay, quantitative RT-PCR (q-RT-PCR), immunoblot, and time-lapse microscopy. We found that five out of six post-treatment cells had higher migration activity than pretreatment cells. However, no unequivocal correlation between increased migration and classic epithelial−mesenchymal transition (EMT) markers could be identified. In fast migrating cells, the microphthalmia-associated transcription factor (MITF) and epidermal growth factor receptor (EGFR) mRNA levels were considerably lower and significantly higher, respectively. Interestingly, high EGFR expression was associated with elevated migration but not with proliferation. Cells with high EGFR expression showed significantly decreased sensitivity to vemurafenib treatment, and had higher Erk activation and FRA-1 expression. Importantly, melanoma cells with higher EGFR expression were more resistant to the EGFR inhibitor erlotinib treatment than cells with lower expression, with respect to both proliferation and migration inhibition. Finally, EGFR-high melanoma cells were characterized by higher PD-L1 expression, which might in turn indicate that immunotherapy may be an effective approach in these cases. |
| first_indexed | 2024-04-12T19:36:58Z |
| format | Article |
| id | doaj.art-dea5f907b1674f4c9a511001df48093a |
| institution | Directory Open Access Journal |
| issn | 1422-0067 |
| language | English |
| last_indexed | 2024-04-12T19:36:58Z |
| publishDate | 2019-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-dea5f907b1674f4c9a511001df48093a2022-12-22T03:19:11ZengMDPI AGInternational Journal of Molecular Sciences1422-00672019-09-012018448410.3390/ijms20184484ijms20184484Long-Term Vemurafenib Exposure Induced Alterations of Cell Phenotypes in Melanoma: Increased Cell Migration and Its Association with EGFR ExpressionEszter Molnár0Tamás Garay1Marco Donia2Marcell Baranyi3Dominika Rittler4Walter Berger5József Tímár6Michael Grusch7Balázs Hegedűs82nd Department of Pathology, Semmelweis University, H-1091 Budapest, HungaryPázmány Péter Catholic University, Faculty of Information Technology and Bionics, H-1083 Budapest, HungaryNational Center for Cancer Immune Therapy, Department of Oncology, Herlev and Gentofte Hospital, University of Copenhagen, DK-2730 Herlev, Denmark2nd Department of Pathology, Semmelweis University, H-1091 Budapest, Hungary2nd Department of Pathology, Semmelweis University, H-1091 Budapest, HungaryInstitute of Cancer Research, Medical University of Vienna, A-1090 Vienna, Austria2nd Department of Pathology, Semmelweis University, H-1091 Budapest, HungaryInstitute of Cancer Research, Medical University of Vienna, A-1090 Vienna, Austria2nd Department of Pathology, Semmelweis University, H-1091 Budapest, HungaryAcquired resistance during BRAF inhibitor therapy remains a major challenge for melanoma treatment. Accordingly, we evaluated the phenotypical and molecular changes of isogeneic human V600E BRAF-mutant melanoma cell line pairs pre- and post-treatment with vemurafenib. Three treatment naïve lines were subjected to in vitro long-term vemurafenib treatment while three pairs were pre- and post-treatment patient-derived lines. Molecular and phenotypical changes were assessed by Sulforhodamine-B (SRB) assay, quantitative RT-PCR (q-RT-PCR), immunoblot, and time-lapse microscopy. We found that five out of six post-treatment cells had higher migration activity than pretreatment cells. However, no unequivocal correlation between increased migration and classic epithelial−mesenchymal transition (EMT) markers could be identified. In fast migrating cells, the microphthalmia-associated transcription factor (MITF) and epidermal growth factor receptor (EGFR) mRNA levels were considerably lower and significantly higher, respectively. Interestingly, high EGFR expression was associated with elevated migration but not with proliferation. Cells with high EGFR expression showed significantly decreased sensitivity to vemurafenib treatment, and had higher Erk activation and FRA-1 expression. Importantly, melanoma cells with higher EGFR expression were more resistant to the EGFR inhibitor erlotinib treatment than cells with lower expression, with respect to both proliferation and migration inhibition. Finally, EGFR-high melanoma cells were characterized by higher PD-L1 expression, which might in turn indicate that immunotherapy may be an effective approach in these cases.https://www.mdpi.com/1422-0067/20/18/4484melanomaV600E BRAF mutationvemurafenib resistanceEGFRPD-L1migration |
| spellingShingle | Eszter Molnár Tamás Garay Marco Donia Marcell Baranyi Dominika Rittler Walter Berger József Tímár Michael Grusch Balázs Hegedűs Long-Term Vemurafenib Exposure Induced Alterations of Cell Phenotypes in Melanoma: Increased Cell Migration and Its Association with EGFR Expression International Journal of Molecular Sciences melanoma V600E BRAF mutation vemurafenib resistance EGFR PD-L1 migration |
| title | Long-Term Vemurafenib Exposure Induced Alterations of Cell Phenotypes in Melanoma: Increased Cell Migration and Its Association with EGFR Expression |
| title_full | Long-Term Vemurafenib Exposure Induced Alterations of Cell Phenotypes in Melanoma: Increased Cell Migration and Its Association with EGFR Expression |
| title_fullStr | Long-Term Vemurafenib Exposure Induced Alterations of Cell Phenotypes in Melanoma: Increased Cell Migration and Its Association with EGFR Expression |
| title_full_unstemmed | Long-Term Vemurafenib Exposure Induced Alterations of Cell Phenotypes in Melanoma: Increased Cell Migration and Its Association with EGFR Expression |
| title_short | Long-Term Vemurafenib Exposure Induced Alterations of Cell Phenotypes in Melanoma: Increased Cell Migration and Its Association with EGFR Expression |
| title_sort | long term vemurafenib exposure induced alterations of cell phenotypes in melanoma increased cell migration and its association with egfr expression |
| topic | melanoma V600E BRAF mutation vemurafenib resistance EGFR PD-L1 migration |
| url | https://www.mdpi.com/1422-0067/20/18/4484 |
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