Treating cells for 24 h with gefitinib effec tively suppressed EGF induced Cox 2 expression in MCF 7/DOX cells. selleck chemical Seliciclib We then tested the effects of the EGFR inhibitor on invasion of MCF 7/ DOX cells. Gefitinib decreased the www.selleckchem.com/products/BAY-73-4506.html invasive potential of MCF 7/DOX cells, indicating that EGFR contributes to invasiveness of selleck chem MCF 7/DOX cells by upregulating Cox 2. Effect of EGFR on PI3K/Akt and MAPK mediated Cox 2 expression in MCF 7/DOX cells Because EGFR controls several other pathways, such as the PI3K/Akt and MAPK pathways, we next investigated which downstream pathway was involved in EGFR mediated Cox 2 expression. As expected, treating MCF 7/DOX cells with EGF for 8 h to induce Cox 2 expression activated both the PI3K/Akt and MAPK pathways.

To confirm the role of the PI3K/ Akt and MAPK pathways in Cox 2 expression, we stu died the effect of the PI3K/Akt inhibitor LY294002 and the MAPK inhibitor U0126 on EGF induced expression of pAkt and Cox 2 in MCF 7/DOX cells. Western blot analysis showed that LY294002 and Inhibitors,Modulators,Libraries U0126 Inhibitors,Modulators,Libraries dramatically Inhibitors,Modulators,Libraries suppressed activation of pAkt and pERK1/2, respec tively, and downregulated EGF induced Cox 2 expres sion. To investigate the role of the PI3K/Akt pathway in invasiveness of MCF 7/DOX cells, we deter mined whether blocking the PI3K/Akt pathway would inhibit invasion of MCF 7/DOX cells in an in vitro inva sion assay. Blocking the PI3K/Akt pathway with LY294002 or Inhibitors,Modulators,Libraries the MAPK Inhibitors,Modulators,Libraries pathway with U0126 dramati cally inhibited invasion Inhibitors,Modulators,Libraries of MCF 7/DOX Inhibitors,Modulators,Libraries cells, but did not affect proliferation of the cells.

Because we wondered whether PI3K/Akt or MAPK sig naling Inhibitors,Modulators,Libraries alone not through Cox 2/PGE2 can increase expression of MMP 2, MMP 9, and uPA, we tested Inhibitors,Modulators,Libraries MMP 2, MMP 9, and uPA mRNA expression Inhibitors,Modulators,Libraries in MCF 7/DOX cells Inhibitors,Modulators,Libraries transfected with siRNA specific for Cox Inhibitors,Modulators,Libraries 2. Transfection with siRNAs targeting Cox 2 specifically inhibited MMP 2, MMP 9, or uPA RNA expression in MCF 7/DOX cells, whereas control scrambled Inhibitors,Modulators,Libraries siRNA demonstrated no effect. Expression of MMP 2, MMP 9 and uPA mRNA was suppressed by the PI3K/Akt inhibitor LY294002 and the MAPK inhibitor U0126 in MCF 7/DOX cells transfected with control siRNA.

however, their expressions were Inhibitors,Modulators,Libraries less affected by either LY294002 or U0126 in MCF 7/DOX cells transfected with Cox 2 siRNA.

selleck catalog Role of EP1 and EP3 receptors in Cox 2 mediated invasion of MCF 7/DOX cells PGE2, which is produced by Cox 2, exerts its effect by binding to specific Inhibitors,Modulators,Libraries prostanoid receptors, which are a family of G protein coupled receptors.

Thus, we investigated the role of EP receptors in the Cox 2 mediated invasion in MCF 7/ DOX cells. First, we compared the basal expression levels of the EP receptors in MCF 7 and MCF 7/DOX www.selleckchem.com/products/Gemcitabine-Hydrochloride(Gemzar).html cells by RT PCR. We found that EP1 and EP3 mRNAs were upregulated, while EP2 and EP4 mRNAs were downregulated in MCF 7/DOX cells. Simi larly, specific induction of EP1 and EP3 selleck kinase inhibitor mRNA, but not EP2 and EP4 mRNA was observed in MCF 7/DOX cells treated with PGE2 for 24 h.