Table 1.
PCR Primer.
Figure 1.
Morphological changes in cells treated with CCL18 and TGFbeta: untreated A549 cells show typical epithelial morphology with close cell-cell-contacts, which are marked by white arrows (A). After treatment with 2 ng/ml -β cells changed the phenotype to a more mesenchymal morphology with prolonged cell extrusions and loosened cell-cell-contacts marked by arrowheads. (B) CCL18 also changes the cell morphology in a dose dependent manner (C [1 ng/ml] and D [10 ng/ml]) to fibroblast-like mesenchymal phenotype and loosened cell-cell-contacts marked by arrowheads.
Figure 2.
mRNA levels of (A) E-cadherin, (B) SNAIL1, (C) FSP-1 after stimulation with -β or various CCL18 concentrations for 72 h relative to the non-stimulated control. Each bar represents means ±SD of four independent experiments. GAPDH was used as housekeeping gene to normalize for differences in the amount of total RNA.
Figure 3.
Effects of CCL18 on the expression of EMT-related proteins (SNAIL1, E-cadherin and FSP-1) after 72 hours of treatment.
All results were normalized to the untreated control. Bars summarize the results of three independent western blot analyses, in the right panel one representative blot is depicted.
Figure 4.
CCL18 induces chemotaxis in A549 lung cancer cells.
Significances were calculated using paired comparison (Wilcoxon Signed Rank Test), control: untreated cells (n = 4).
Figure 5.
CCL 18 induces invasion of A549 lung cancer cells into a extracellular matrix derived from the Engelbreth Holm-Swarm (EHS) mouse tumor.
Significances were calculated using paired comparison (Wilcoxon Signed Rank Test), control: untreated cells (n = 3).
Figure 6.
CCL18 suppreses proliferation of A549 cells. (n = 7 in each group).
Figure 7.
Influence of CLL18 on chemoresistance.
Cells were cultured in presence or absence of -TGFbeta or CCL18 in the concentrations indicated for 72 h. After the culture period, medium was changed and the cells were incubated with cisplatin for additional 72 hours at concentrations ranging from 0.4 to 25 µg/ml. Cell survival was measured by MTT assay. The horizontal line indicates the LD50, vertical lines indicate the LD50 for the respective pre-incubation condition (n = 4).