Simultaneous blocking of IL-6 and IL-8 is sufficient to fully inhibit CAF-induced human melanoma cell invasiveness.
Tumour microenvironment plays a critical role in cell invasion and metastasis. To investigate the role of cancer-associated fibroblasts (CAFs) in melanoma cell invasiveness, we used 3D spheroid invasion assay. The effect of conditioned media from normal fibroblasts and CAFs cultivated alone or co-cultivated with melanoma cells on BLM or A2058 melanoma spheroid invasion was analysed. We found that conditioned media from CAFs and CAFs co-cultured with melanoma cells, especially, promote invasion and migration, without significant effect on melanoma cell proliferation. We further analysed the expression of pro-invasive cytokines IL-8 and IL-6 in media and found that melanoma cells are dominant producers of IL-8 and fibroblasts are dominant producers of IL-6 in 2D monocultures, while co-cultivation of CAFs with melanoma cells induces production/secretion of IL-6 and IL-8 into the media. The analyses of IL-6 levels in 3D cultures and human melanoma samples, however, revealed that at least in some cases IL-6 is also produced directly by melanoma cells. Analysis of the role of IL-6 and IL-8 in CAF-induced melanoma invasion, using neutralising antibodies, revealed that simultaneous blocking of IL-6 and IL-8 is sufficient to fully inhibit CAF-induced human melanoma cell invasiveness. In summary, these experiments indicate the important role of CAFs and IL-8 and IL-6 cytokines in melanoma cell invasiveness.
Department of Cell Biology, Charles University in Prague, Viničná 7, Prague, Czech Republic.|Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University (BIOCEV), Průmyslová 595, 25242, Vestec u Prahy, Czech Republic.
Njainday Pulo
NP
Jobe
Department of Cell Biology, Charles University in Prague, Viničná 7, Prague, Czech Republic.|Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University (BIOCEV), Průmyslová 595, 25242, Vestec u Prahy, Czech Republic.
Daniel
D
Rösel
1st Faculty of Medicine, Institute of Anatomy, Charles University, U Nemocnice 3, 12800, Prague 2, Czech Republic.|Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University (BIOCEV), Průmyslová 595, 25242, Vestec u Prahy, Czech Republic.
Barbora
B
Dvořánková
1st Faculty of Medicine, Institute of Anatomy, Charles University, U Nemocnice 3, 12800, Prague 2, Czech Republic.|Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University (BIOCEV), Průmyslová 595, 25242, Vestec u Prahy, Czech Republic.|Department of Dermatovenerology, 1st Faculty of Medicine, Charles University and General University Hospital, U Nemocnice 2, 12808, Prague 2, Czech Republic.
Ondřej
O
Kodet
1st Faculty of Medicine, Institute of Anatomy, Charles University, U Nemocnice 3, 12800, Prague 2, Czech Republic.|Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University (BIOCEV), Průmyslová 595, 25242, Vestec u Prahy, Czech Republic.
Lukáš
L
Lacina
1st Faculty of Medicine, Institute of Anatomy, Charles University, U Nemocnice 3, 12800, Prague 2, Czech Republic.
Rosana
R
Mateu
1st Faculty of Medicine, Institute of Anatomy, Charles University, U Nemocnice 3, 12800, Prague 2, Czech Republic. karel.smetana@lf1.cuni.cz.
Karel
K
Smetana
Department of Cell Biology, Charles University in Prague, Viničná 7, Prague, Czech Republic. brabek@natur.cuni.cz.|Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University (BIOCEV), Průmyslová 595, 25242, Vestec u Prahy, Czech Republic. brabek@natur.cuni.cz.
Jan
J
Brábek
D000072645:Cancer-Associated Fibroblasts / Q000187:drug effects* / Q000473:pathology; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007150:Immunohistochemistry; D015850:Interleukin-6 / Q000032:analysis / Q000378:metabolism; D016209:Interleukin-8 / Q000032:analysis / Q000037:antagonists & inhibitors* / Q000378:metabolism; D008545:Melanoma / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology*; D009361:Neoplasm Invasiveness / Q000517:prevention & control*; D000093242:Interleukin-6 Inhibitors
D017077:Culture Media, Conditioned; D015850:Interleukin-6; D016209:Interleukin-8; D000093242:Interleukin-6 Inhibitors
3D culture of melanoma cells; Cancer microenvironment; Cancer-associated fibroblasts; Chemokine; Cytokine; Invasiveness