Journal:

Article Title: Interleukin-10 promotes B16-melanoma growth by inhibition of macrophage functions and induction of tumour and vascular cell proliferation

doi: 10.1046/j.1365-2567.2002.01363.x

Figure Lengend Snippet: Concentration and activity of IL-10 secreted by IL-10-transfected B16-melanoma cell lines. The B16-melanoma cell line was transfected with IL-10 cDNA and the production of IL-10 was quantified from cell culture supernatants by ELISA. Three clones that secreted high (B16-10), medium (B16-13) and low (B16-5) amounts of IL-10 were selected. Results are expressed in pg/ml of IL-10/2 × 106 cells/48 hr. The biological activity of the secreted IL-10 was determined by measuring the nitrite (NO2) concentrations after macrophages were incubated with cell line supernatants and then activated by LPS/IFN-γ (see Material and Methods). Labels at the left of the slash indicate the cell-line supernatant and the sign ‘−’ indicates not incubated with cell-line supernatant; ‘+’ or ‘−’ signals at the right of the slash indicate activation or not activation of macrophages. The bar representing the nitrite basal production by macrophages is indicated as ‘−/−’ (no cell line supernatant/not activated) and total nitrite production after stimulation is indicated as −/+ (no cell line supernatant/activated). The mean of triplicates is presented and error bars represent the standard errors of means.

Article Snippet: Then they were stimulated with 10 ng LPS (Sigma Chemical Co.) and 100 ng IFN-γ (Schering-Plough, Mexico City, Mexico) in 500 µl RPMI-1640 for 48 hr.

Techniques: Concentration Assay, Activity Assay, Transfection, Cell Culture, Enzyme-linked Immunosorbent Assay, Clone Assay, Incubation, Activation Assay

Journal:

Article Title: Interleukin-10 promotes B16-melanoma growth by inhibition of macrophage functions and induction of tumour and vascular cell proliferation

doi: 10.1046/j.1365-2567.2002.01363.x

Figure Lengend Snippet: Effects of IL-10 on the interactions of immune and tumour cells in vitro

Article Snippet: Then they were stimulated with 10 ng LPS (Sigma Chemical Co.) and 100 ng IFN-γ (Schering-Plough, Mexico City, Mexico) in 500 µl RPMI-1640 for 48 hr.

Techniques: Lysis, Expressing

Journal: bioRxiv

Article Title: Deficiency of ZC3HC1 increases vascular smooth muscle cell migration, proliferation and neointima formation following injury

doi: 10.1101/2021.09.29.462212

Figure Lengend Snippet: A) Predominant association of rs11556924 with cardiovascular disease (CVD) driven by vascular SMC dysfunction. The plot shows the beta effect size of the rs11556924-T allele derived from several genome-wide association studies (GWAS) categorized according to eight CVD-related traits (BP, blood pressure; CAD, coronary artery disease; MI, myocardial infarction; cIMT, carotid intima-media thickness; chol, cholesterol; WBC, white blood cell count; PC, platelet count). Asterisks indicate genome-wide significance (p<5 × 10 -8 ); data points without asterisks represent genome-wide associations with suggestive significance (p<5 × 10 -5 ). Negative effect size indicates that the risk allele (T) of the single nucleotide polymorphism, rs11556924, is associated with a lower risk for several traits, including blood pressure or coronary artery disease. B) The risk allele (T) of rs11556924 is associated with lower ZC3HC1 expression and C) faster migration by vascular SMCs. SMCs transfected with siRNA against ZC3HC1 showed significant D) ZC3HC1 downregulation and E) increased migration in the presence or absence of PDGF-BB (n=3, eight images per replicate). F) Representative images of the migration of SMCs transfected with ZC3HC1 siRNA and control siRNA (scale bars, 100 μm). G) Knockdown of ZC3HC1 resulted in significant increases in cell proliferation in the presence of PDGF-BB. Values are shown as means. The translucent error bands represent s.d. * p<0.05; ** p<0.01 ; n.s., not significant. Data in B , C , and E were analyzed using unpaired Student’s t test. Others were analyzed using Mann-Whitney U test.

Article Snippet: These human SMCs were subsequently treated with 100 ng/mL PDGF-BB (Peprotech) to induce proliferation.

Techniques: Derivative Assay, GWAS, Cell Counting, Genome Wide, Expressing, Migration, Transfection, MANN-WHITNEY

Journal: bioRxiv

Article Title: Deficiency of ZC3HC1 increases vascular smooth muscle cell migration, proliferation and neointima formation following injury

doi: 10.1101/2021.09.29.462212

Figure Lengend Snippet: A) Volcano plot of the expression profiles of differentially expressed genes in SMCs transfected with siRNA against ZC3HC1 (ZC3HC1 siRNA) (n=4) and control siRNA (n=5). The red data points represent the differentially expressed genes with statistical significance, whereas the gray data points indicate genes without disturbed gene expression. The vertical dashed lines correspond to a 0.5-fold-change in gene expression (up or down), and the horizontal dashed line represents the adjusted p-value for each gene. B) Heat map showing contractile SMC marker genes that were differentially expressed upon knockdown of ZC3HC1 (from the same batch) in the presence or absence of PDGF-BB. C) qPCR validation of the expression of the contractile SMC marker genes. Values are shown as mean ± s.d.; * p<0.05; n.s., not significant. Data in C were analyzed using Mann-Whitney U test.

Article Snippet: These human SMCs were subsequently treated with 100 ng/mL PDGF-BB (Peprotech) to induce proliferation.

Techniques: Expressing, Transfection, Marker, MANN-WHITNEY

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