Simultaneous blocking of IL-6 and IL-8 is sufficient to fully inhibit CAF-induced human melanoma cell invasiveness
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Published on: October 20, 2016
Njainday Pulo Jobe1,2, Daniel Rösel1,2, Barbora Dvořánková3,2
1Department of Cell Biology, Charles University in Prague, Viničná 7, Prague, Czech Republic.
Abstract:
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.
Insights
Cancer-associated fibroblasts (CAFs) promote melanoma cell invasion and migration. Blocking IL-6 and IL-8 cytokines fully inhibits this CAF-induced invasiveness, highlighting their critical role in melanoma metastasis.
Area of Science:
- Oncology
- Cell Biology
- Cancer Research
Background:
- Tumor microenvironment significantly influences cancer cell invasion and metastasis.
- Cancer-associated fibroblasts (CAFs) are key components of the tumor microenvironment.
- Understanding the interaction between CAFs and melanoma cells is crucial for developing targeted therapies.
Purpose of the Study:
- To investigate the role of cancer-associated fibroblasts (CAFs) in promoting melanoma cell invasiveness.
- To identify the specific cytokines involved in CAF-mediated melanoma cell invasion.
- To evaluate the therapeutic potential of blocking key cytokines in inhibiting melanoma cell invasion.
Main Methods:
- Utilized a 3D spheroid invasion assay to model melanoma cell invasion.
- Analyzed conditioned media from normal fibroblasts and CAFs, alone and co-cultured with melanoma cells.
- Quantified the expression of pro-invasive cytokines IL-6 and IL-8.
- Employed neutralising antibodies to block IL-6 and IL-8 activity in co-culture models.
Main Results:
- Conditioned media from CAFs, especially when co-cultured with melanoma cells, significantly enhanced melanoma cell invasion and migration.
- Melanoma cells were primary producers of IL-8, while fibroblasts produced IL-6 in monocultures.
- Co-cultivation induced increased secretion of both IL-6 and IL-8.
- Simultaneous blockade of IL-6 and IL-8 completely inhibited CAF-induced melanoma cell invasiveness.
Conclusions:
- Cancer-associated fibroblasts play a significant role in promoting melanoma cell invasion and migration.
- Interleukin-6 (IL-6) and Interleukin-8 (IL-8) are critical mediators of CAF-induced melanoma invasiveness.
- Targeting both IL-6 and IL-8 presents a promising therapeutic strategy to inhibit melanoma metastasis.
