Coffee polyphenols change the expression of STAT5B and ATF-2 modifying cyclin D1 levels in cancer cells - PubMed (original) (raw)
Coffee polyphenols change the expression of STAT5B and ATF-2 modifying cyclin D1 levels in cancer cells
Carlota Oleaga et al. Oxid Med Cell Longev. 2012.
Abstract
Background: Epidemiological studies suggest that coffee consumption reduces the risk of cancer, but the molecular mechanisms of its chemopreventive effects remain unknown.
Objective: To identify differentially expressed genes upon incubation of HT29 colon cancer cells with instant caffeinated coffee (ICC) or caffeic acid (CA) using whole-genome microarrays.
Results: ICC incubation of HT29 cells caused the overexpression of 57 genes and the underexpression of 161, while CA incubation induced the overexpression of 12 genes and the underexpression of 32. Using Venn-Diagrams, we built a list of five overexpressed genes and twelve underexpressed genes in common between the two experimental conditions. This list was used to generate a biological association network in which STAT5B and ATF-2 appeared as highly interconnected nodes. STAT5B overexpression was confirmed at the mRNA and protein levels. For ATF-2, the changes in mRNA levels were confirmed for both ICC and CA, whereas the decrease in protein levels was only observed in CA-treated cells. The levels of cyclin D1, a target gene for both STAT5B and ATF-2, were downregulated by CA in colon cancer cells and by ICC and CA in breast cancer cells.
Conclusions: Coffee polyphenols are able to affect cyclin D1 expression in cancer cells through the modulation of STAT5B and ATF-2.
Figures
Figure 1
Biological association network (BAN) of differentially expressed genes in common between CA and ICC. The list of common genes between both treatments was used to construct a BAN with the Pathway Analysis software within GeneSpring v.11.5.1. An expanded network was constructed by setting an advanced filter that included the categories of binding, expression, metabolism, promoter binding, protein modification, and regulation. Only proteins are represented. The BAN shows the node genes STAT5B and ATF-2 that were further studied.
Figure 2
Quantitation of mRNA and protein levels for STAT5B and ATF-2 in HT29 cells. The mRNA levels of STAT5B (a) and ATF-2 (b) were determined in control HT29 cells (empty bars) and cells treated with caffeic acid (CA, filled bars) and instant caffeinated coffee (ICC, grey bars) by RT real-time PCR as described in Methods. Results are expressed in fold changes compared to the control and are the mean ± SE of 3 different experiments. *P < 0.05 compared with the corresponding control. The protein levels of STAT5B (c) and ATF-2 (d) were determined in control HT29 cells (empty bars) and cells treated with caffeic acid (CA, filled bars) and instant caffeinated coffee (ICC, grey bars) by Western blot. Blots were reprobed with an antibody against _β_-actin or tubulin to normalize the results. Results represent the mean ± SE of 3 different experiments. *P < 0.05 and **P < 0.01 compared with the corresponding control.
Figure 3
Expression of cyclin D1 upon incubation with ICC and CA in HT29 and MCF-7 cells. (a) Quantitation of STAT5b (empty bars), ATF-2 (filled bars), and cyclin D1 (grey bars) protein levels in MCF-7 cells. The protein levels were determined in control MCF-7 cells (CNT) and cells treated with caffeic acid (CA) and instant coffee (ICC) by Western blot. Blots were reprobed with an antibody against _β_-actin to normalize the results. Results represent the mean ± SE of 3 different experiments. *P < 0.05 and ***P < 0.001 compared with the corresponding control. (b) Quantitation of STAT5b (empty bars), ATF-2 (filled bars), and cyclin D1 (grey bars) protein levels in HT29 cells. The protein levels were determined in control HT29 cells (CNT) and cells treated with caffeic acid (CA) and instant coffee (ICC) by Western blot. Blots were reprobed with an antibody against _β_-actin to normalize the results. Results represent the mean ± SE of 3 different experiments.*P < 0.05 and **P < 0.01 compared with the corresponding control.
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