Open Access

Microenvironment‑driven resistance to B‑Raf inhibition in a melanoma patient is accompanied by broad changes of gene methylation and expression in distal fibroblasts

  • Authors:
    • Ondřej Kodet
    • Barbora Dvořánková
    • Běla Bendlová
    • Vlasta Sýkorová
    • Ivana Krajsová
    • Jiří Štork
    • Jan Kučera
    • Pavol Szabo
    • Hynek Strnad
    • Michal Kolář
    • Čestmír Vlček
    • Karel Smetana
    • Lukáš Lacina
  • View Affiliations

  • Published online on: February 1, 2018     https://doi.org/10.3892/ijmm.2018.3448
  • Pages: 2687-2703
  • Copyright: © Kodet et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The incidence of malignant melanoma is rapidly increasing and current medicine is offering only limited options for treatment of the advanced disease. For B‑Raf mutated melanomas, treatment with mutation‑specific drug inhibitors may be used. Unfortunately, tumors frequently acquire resistance to the treatment. Tumor microenvironment, namely cancer‑associated fibroblasts, largely influence this acquired resistance. In the present study, fibroblasts were isolated from a patient suffering from acrolentiginous melanoma (Breslow, 4.0 mm; Clark, IV; B‑Raf V600E mutated). The present study focused on the expression of structural and functional markers of fibroblast activation in melanoma‑associated fibroblasts (MAFs; isolated prior to therapy initiation) as well as in autologous control fibroblasts (ACFs) of the same patient isolated during B‑Raf inhibitor therapy, yet before clinical progression of the disease. Analysis of gene transcription was also performed, as well as DNA methylation status analysis at the genomic scale of both isolates. MAFs were positive for smooth muscle actin (SMA), which is a marker of myofibroblasts and the hallmark of cancer stoma. Surprisingly, ACF isolated from the distant uninvolved skin of the same patient also exhibited strong SMA expression. A similar phenotype was also observed in control dermal fibroblasts (CDFs; from different donors) exclusively following stimulation by transforming growth factor (TGF)‑β1. Immunohistochemistry confirmed that melanoma cells potently produce TGF‑β1. Significant differences were also identified in gene transcription and in DNA methylation status at the genomic scale. Upregulation of SMA was observed in ACF cells at the protein and transcriptional levels. The present results support recent experimental findings that tumor microenvironment is driving resistance to B‑Raf inhibition in patients with melanoma. Such an activated microenvironment may be viable for the growth of circulating melanoma cells.
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May-2018
Volume 41 Issue 5

Print ISSN: 1107-3756
Online ISSN:1791-244X

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Spandidos Publications style
Kodet O, Dvořánková B, Bendlová B, Sýkorová V, Krajsová I, Štork J, Kučera J, Szabo P, Strnad H, Kolář M, Kolář M, et al: Microenvironment‑driven resistance to B‑Raf inhibition in a melanoma patient is accompanied by broad changes of gene methylation and expression in distal fibroblasts. Int J Mol Med 41: 2687-2703, 2018
APA
Kodet, O., Dvořánková, B., Bendlová, B., Sýkorová, V., Krajsová, I., Štork, J. ... Lacina, L. (2018). Microenvironment‑driven resistance to B‑Raf inhibition in a melanoma patient is accompanied by broad changes of gene methylation and expression in distal fibroblasts. International Journal of Molecular Medicine, 41, 2687-2703. https://doi.org/10.3892/ijmm.2018.3448
MLA
Kodet, O., Dvořánková, B., Bendlová, B., Sýkorová, V., Krajsová, I., Štork, J., Kučera, J., Szabo, P., Strnad, H., Kolář, M., Vlček, Č., Smetana, K., Lacina, L."Microenvironment‑driven resistance to B‑Raf inhibition in a melanoma patient is accompanied by broad changes of gene methylation and expression in distal fibroblasts". International Journal of Molecular Medicine 41.5 (2018): 2687-2703.
Chicago
Kodet, O., Dvořánková, B., Bendlová, B., Sýkorová, V., Krajsová, I., Štork, J., Kučera, J., Szabo, P., Strnad, H., Kolář, M., Vlček, Č., Smetana, K., Lacina, L."Microenvironment‑driven resistance to B‑Raf inhibition in a melanoma patient is accompanied by broad changes of gene methylation and expression in distal fibroblasts". International Journal of Molecular Medicine 41, no. 5 (2018): 2687-2703. https://doi.org/10.3892/ijmm.2018.3448