Figure 1.
VEGF-A protein expression in C3842 chondrosarcoma cells after treatment with IL-1β (10 ng/ml) for 1 to 24 h, in untreated cells (lane 2) and in VEGF-A overexpressing N109 renal carcinoma cells (lane 1), which served as positive control (P).
β-actin is used as loading control.
Figure 2.
Phosphorylation of IκBα in C3842 (A) and SW1353 (B) chondrosarcoma cells after treatment with IL-1β.
The signals disappeared after 15κBα. In untreated cells IκBα is not phosphorylated (lane 1).
Figure 3.
The time of the incubation with Curcumin prior to IL-1β treatment is essential for the inhibitory effect of Curcumin in chondrosarcoma cells.
After an incubation of at least 60-1β treatment, Curcumin blocks phosphorylation of IκBα in C3842 chondrosarcoma cells.
Figure 4.
An appropriate concentration of Curcumin is necessary for the inhibitory effect on IL-1 signaling.
At least a Curcumin concentration of 15 µmol/l is required to block IκBα phosphorylation in C3842 (A) and SW1353 (B) chondrosarcoma cells.
Figure 5.
Incubation with Curcumin (20 µmol/l) for 120 min blocks IL-1β-induced phosphorylation of IκBα in C3842 and SW1353 chondrosarcoma cells.
Controls with untreated cells, and cells treated either with IL-1β or Curcumin were included.
Figure 6.
Detection of IL-1β-induced nuclear translocation of NF-κB by immunofluorescence in C3842 cells (A–D) and SW1353 cells (E-H).
In untreated cells and cells treated with Curcumin or Curcumin+IL-1β, NF-κB was detected in the cytoplasm, not in the nucleus (Figure 6). In cells treated with IL-1β nuclear translocation of NF-κB was evident.
Figure 7.
Effect of Curcumin on VEGF-A expression.
IL-1β-induced VEGF-A expression is blocked in C3842 and SW1353 chondrosarcoma cells after incubation with Curcumin.
Figure 8.
Angiogenesis assay with quantitative image analysis.
Cell culture supernatants from C3842 cells treated with IL-1β led to increased number and length of microvessel segments (D). Treatment with Curcumin blocked IL-1β-induced tube formation (F). Controls with supernatants from untreated cells (C) and cells treated with Curcumin (E) were included. Control with FCS-free medium (A). Control with medium supplemented with FCCS (B).
Table 1.
Relative expression of genes associated with NF-κB. RNA was extracted from C3842 cells treated with IL-1β (10 ng/ml), Curcumin (20 µmol/l) or both.
Figure 9.
Illustration of the data presented in table 1, representing the number of genes induced (green) or repressed (red) by treatment with IL-1β, Curcumin or IL-1β+Curcumin.
The size of the circles is proportional to the number of genes in each group. The threshold was set over 2 and under 0.5-fold variation in gene expression. Additionally, the Curcumin effect indicating reduced expression is indicated. Numbers in overlaps indicate the number of genes that shared in the groups.