Journal: Frontiers in Immunology

Article Title: IL-1β, IL-8, and Matrix Metalloproteinases-1, -2, and -10 Are Enriched upon Monocyte–Breast Cancer Cell Cocultivation in a Matrigel-Based Three-Dimensional System

doi: 10.3389/fimmu.2017.00205

Figure Lengend Snippet: Primary breast cancer (BrC) cells basal levels of secretion of inflammatory cytokines . (A) The levels of the cytokines, chemokines, and growth factors of interest were determined in the supernatants of the 3D cultures of BrC primary isolates. Each point represents the mean of three determinations made for three independent cultures. The horizontal line represents the average of the primary cultures readings. (B) Comparison of the migratory properties of monocytes in response to GM-CSF, MCP-1, and RANTES. (C) Invasion assay with the commercial BrC cell lines in response to IL-8. Three independent experiments were performed. Statistical significance is indicated by *** p < 0.001 and ** p < 0.01.

Article Snippet: The Transwells were placed in a 24-well culture dish containing 800 μL of their respective media with sera or without sera but supplemented with 100 ng/mL of IL-8 as chemoattractant (PeproTech, Rocky Hill, NJ, USA).

Techniques: Invasion Assay

Journal: Frontiers in Immunology

Article Title: IL-1β, IL-8, and Matrix Metalloproteinases-1, -2, and -10 Are Enriched upon Monocyte–Breast Cancer Cell Cocultivation in a Matrigel-Based Three-Dimensional System

doi: 10.3389/fimmu.2017.00205

Figure Lengend Snippet: Levels of inflammatory cytokines present in supernatants of commercial breast cancer (BrC) cell lines in co-culture with monocytes . Inflammatory cytokines were determined in three-dimensional (3D) individual cultures of the BrC cell lines (indicated as 3D) and in co-culture with the three different sources of monocytes independently [THP-1, U937, and primary monocytes (PM); indicated as cells co-cultured (CC)]. (A) The means and SD of three independent co-culture experiments are shown (in picogram/milliliter). (B) Summary of the cytokines that gave statistically significant differences. For the statistical analysis, MCF-7 and T47D cells were grouped as non-aggressive BrC cells and MDA-MB-231 and HS578T were grouped as aggressive BrC cells . THP-1, U937, and PM were grouped as Monocytes , and statistics were calculated using all combinations of aggressive vs non-aggressive BrC cells with all the types of monocytes. Left panel [interleukin (IL)-1β]: statistical differences were found between BrC cells cultured individually vs in co-culture with monocytes, and between co-cultures of the non-aggressive BrC cells vs the co-cultures of the aggressive cells. Right panel (IL-8): statistical differences were found between non-aggressive cells cultured individually (3D) and the rest of the groups: non-aggressive CC with monocytes and with aggressive cells either cultured individually or in co-culture with monocytes. Statistical significance p ≤ 0.05 was indicated with *, p ≤ 0.01 with **, and p ≤ 0.001 with ***. The statistical tests comparing monocytes against co-cultures showed significant differences only for HA-BrC (IL-1β p ≤ 0.0001 and IL-8 p = 0.0014) but not for NA-BrC cells (IL-1β p = 0.752 and IL-8 p = 0.0942).

Article Snippet: The Transwells were placed in a 24-well culture dish containing 800 μL of their respective media with sera or without sera but supplemented with 100 ng/mL of IL-8 as chemoattractant (PeproTech, Rocky Hill, NJ, USA).

Techniques: Co-Culture Assay, Cell Culture

Journal: Frontiers in Immunology

Article Title: IL-1β, IL-8, and Matrix Metalloproteinases-1, -2, and -10 Are Enriched upon Monocyte–Breast Cancer Cell Cocultivation in a Matrigel-Based Three-Dimensional System

doi: 10.3389/fimmu.2017.00205

Figure Lengend Snippet: Levels of interleukin (IL)-1β and IL-8 increased when primary breast cancer (BrC) cells were co-cultured with monocytes . Levels of IL-1β (left panel) and IL-8 (right panel) significantly increased when primary cells derived from the eight BrC patients were co-cultured with the primary monocytes from healthy donors. Three independent experiments were carried out and each point represents the mean of all determinations for each BrC primary isolate. Statistical significance p ≤ 0.05 is indicated with * and p ≤ 0.01 with **.

Article Snippet: The Transwells were placed in a 24-well culture dish containing 800 μL of their respective media with sera or without sera but supplemented with 100 ng/mL of IL-8 as chemoattractant (PeproTech, Rocky Hill, NJ, USA).

Techniques: Cell Culture, Derivative Assay

Journal: Frontiers in Immunology

Article Title: IL-1β, IL-8, and Matrix Metalloproteinases-1, -2, and -10 Are Enriched upon Monocyte–Breast Cancer Cell Cocultivation in a Matrigel-Based Three-Dimensional System

doi: 10.3389/fimmu.2017.00205

Figure Lengend Snippet: Working model . Tumor cells coerce monocytes/macrophages to fulfill activities more attune with tumor grow. MCP-1, G-SCF, and RANTES may be critical components of the message send by tumor cells. Monocytes/macrophages respond helping to establish an inflammatory microenvironment rich in cytokines with pro-tumor activities such as interleukin (IL)-1β and IL-8, which paradoxically may be immunosuppressive of cytotoxic T cell function, since it is high in the previously mentioned cytokines plus GM-CSF and low in INF-α2 and IL-12p70. This inflammatory microenvironment is also enriched of metalloproteinases (MMPs) that promote the degradation of ECM. Altogether, this microenvironment may facilitate the epithelial to mesenchymal transition (EMT), which facilitates invasion and probably metastasis and chemoresistance. The most aggressive tumor cells seem to be more efficient to establish this pro-tumoral microenvironment that further boosts tumor aggressiveness, thus creating a positive feedback loop.

Article Snippet: The Transwells were placed in a 24-well culture dish containing 800 μL of their respective media with sera or without sera but supplemented with 100 ng/mL of IL-8 as chemoattractant (PeproTech, Rocky Hill, NJ, USA).

Techniques: Cell Function Assay