S100A7 (Psoriasin), highly expressed in Ductal Carcinoma (DCIS), is regulated by IFN-gamma in mammary epithelial cells-4

<p><b>Copyright information:</b></p><p>Taken from "S100A7 (Psoriasin), highly expressed in Ductal Carcinoma (DCIS), is regulated by IFN-gamma in mammary epithelial cells"</p><p>http://www.biomedcentral.com/1471-2407/7/205</p><p>BMC Cancer 2007;7():205-205.</p><p>Published online 6 Nov 2007</p><p>PMCID:PMC2180183.</p><p></p>on with dnIkkβ-virus in MCF10A cells. Neither infection with the different adenoviruses nor the IFN-gamma had any effect on the protein levels of pro-caspase-3. The function of the virus was verified by the accumulation of the IκBα protein. Tubulin assesses equal loading. C control

S100A7 (Psoriasin), highly expressed in Ductal Carcinoma (DCIS), is regulated by IFN-gamma in mammary epithelial cells-1

<p><b>Copyright information:</b></p><p>Taken from "S100A7 (Psoriasin), highly expressed in Ductal Carcinoma (DCIS), is regulated by IFN-gamma in mammary epithelial cells"</p><p>http://www.biomedcentral.com/1471-2407/7/205</p><p>BMC Cancer 2007;7():205-205.</p><p>Published online 6 Nov 2007</p><p>PMCID:PMC2180183.</p><p></p> reaction (RT-PCR), mRNA level of psoriasin was downregulated in response to IFN-gamma in MCF10A cells () and in MDA-MB-468 cells (). Expression data for psoriasin are presented as ratios, in which the expression data are normalized to an endogenous control (β-actin). Values obtained in control cells were designed as 1 and values obtained from IFN-gamma treated cells were normalized to this from the same run. The data are presented as the mean of three different runs

S100A7 (Psoriasin), highly expressed in Ductal Carcinoma (DCIS), is regulated by IFN-gamma in mammary epithelial cells-3

<p><b>Copyright information:</b></p><p>Taken from "S100A7 (Psoriasin), highly expressed in Ductal Carcinoma (DCIS), is regulated by IFN-gamma in mammary epithelial cells"</p><p>http://www.biomedcentral.com/1471-2407/7/205</p><p>BMC Cancer 2007;7():205-205.</p><p>Published online 6 Nov 2007</p><p>PMCID:PMC2180183.</p><p></p>MTS-assay on days 3 and 7 and the IFN-gamma/control quotient is demonstrated. Treatment with IFN-gamma produced a 59.5% (day 3) and 32% (day 7) reduction in viability as compared to control cells. The assay was performed in duplicate

S100A7 (Psoriasin), highly expressed in Ductal Carcinoma (DCIS), is regulated by IFN-gamma in mammary epithelial cells-6

<p><b>Copyright information:</b></p><p>Taken from "S100A7 (Psoriasin), highly expressed in Ductal Carcinoma (DCIS), is regulated by IFN-gamma in mammary epithelial cells"</p><p>http://www.biomedcentral.com/1471-2407/7/205</p><p>BMC Cancer 2007;7():205-205.</p><p>Published online 6 Nov 2007</p><p>PMCID:PMC2180183.</p><p></p>00 u or 1000 u of IFN-gamma. IFN-gamma treatment had no effect on the protein levels of pro-caspase-3. Calgranulin B and calgranulin A () are not influenced by the IFN-gamma treatment. Equal amounts of protein lysate were loaded on the gel. , Treatment with 75 μM HOfor one hour followed by culture for 48 h in regular medium induces the expression of calgranulin A in MCF10A cells. 100 μg of protein lysate were loaded on the gel. , MDA-MB-468 cells with the endogenous expression of psoriasin show the time-dependent repression of psoriasin expression when treated 24, 48 and 72 hours with 1000 u of IFN-gamma. Equal amounts of protein lysate were loaded on the gel. Probing with tubulin/GAPDH assesses the equal loading of the samples. C = cells cultured in monolayer

Confluence- and suspension-cultured mammary epithelial cells demonstrate increased psoriasin and CD24 expression.

<p>MCF10A cells were cultured in confluence for 5 and 10 days or in suspension for 3 days. <b>A</b> Psoriasin expression, analyzed by Western blotting, was induced in MCF10A cells cultured in confluence and suspension, compared with exponentially growing cells. Equal loading was confirmed by GAPDH. The figure illustrates a representative example (n = 3). <b>B</b> In confluence- and suspension-cultured cells, psoriasin and CD24 expression were increased, whereas CD44 expression was decreased compared with exponentially growing cells. The expression level of CD24 and CD44 was measured using flow cytometry and the expression level of psoriasin was quantified from Western blots. The data are presented as the mean ± SD of relative expression (n = 3). The p-values (*<0.05, **<0.01, ***<0.001) were calculated using the Student's t-test.</p

Inhibition of ROS and the NF-κB signaling pathway suppresses psoriasin and CD24 expression.

<p>MCF10A cells were cultured in confluence for 10 days. The inhibition of ROS using NAC (<b>A</b>) and NF-κB using CAPE (<b>B</b>) and dnIKKB (<b>C</b>) led to a significantly reduction in CD24 and psoriasin expression in confluence-cultured MCF10A, measured by flow cytometry and Western blot, respectively. No reduction of CD24 expression was seen by the inhibition of PLC-IP3 using U73122 (<b>D</b>). The reduced CD24 expression by Tyrphostin (<b>E</b>) did not reach statistical significance. The inhibition of PI3-K by Wortmannin (<b>F</b>) showed no decrease in psoriasin or CD24 expression. The inhibitors showed no effect on CD44 expression, except for treatment with NAC (<b>A</b>), which increased the expression of CD44. Western blot inserts illustrate representative examples (n = 4) of the psoriasin expression. Equal loading was confirmed by GAPDH. The data are presented as the mean ± SD of relative expression (n = 4). The p-values (*<0.05, **<0.01, ***<0.001) were calculated using the Student's t-test.</p

Psoriasin and MUC1 expression is elevated in CD24⁺ mammary epithelial cells.

<p>MCF10A cells were cultured in confluence for 10 days and separated for CD24⁺ cells, using magnetic activated cell sorting. <b>A</b> Psoriasin expression, analyzed by Western blotting, was confined to the CD24⁺ cell fraction, compared with negative selection and controls. Equal loading was confirmed by GAPDH. The figure illustrates a representative example (n = 3). <b>B</b> Separated CD24⁺ cells showed an increase in the expression of CD24 and a decrease in the expression of CD44, compared with negative selection. The expression of MUC1 was increased in the same level as CD24 expression. The figures illustrate representative examples (n = 4).</p

Endogenous psoriasin causes increased CD24 expression.

<p>MCF10A cells were transfected with a psoriasin-targeting shRNA (Pso-shRNA) or siRNA (Pso-siRNA), and their corresponding controls,(C-shRNA and C-siRNA). Transfected MCF10A cells were cultured in confluence for 10 days or in suspension for 3 days. The expression level of psoriasin was detected by Western blot. CD24 and CD44 expression were measured using flow cytometry. <b>A</b> No induction of psoriasin expression was observed in Pso-shRNA, compared with C-shRNA, in confluence or suspension. <b>B</b> Pso-shRNA showed a decrease in the expression of CD24 and an increase in the expression of CD44, compared with C-shRNA, during confluence and suspension. <b>C</b> The expression of psoriasin in Pso-siRNA was dramatically downregulated, compared with C-siRNA. <b>D</b> Pso-siRNA in suspension culture showed a reduced CD24 expression, compared with C-siRNA. Equal loading was confirmed by GAPDH. The figures illustrate representative examples (n = 3). The data are presented as the mean ± SD of relative expression (n = 3). The p-values (*<0.05) were calculated using the Student's t-test.</p