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ibm proprinter x24 windows 7 downloadIn situ measurement of cyclo-oxygenase-2 expression and prostaglandin biosynthesis by flow cytometry.
The expression of cyclo-oxygenase-2 (COX-2) and subsequent PGE2 synthesis can be induced by different factors, including certain cytokines. Prostaglandins are important mediators in cellular function and influence growth, proliferation, and migration. Therefore, PGE2 synthesis may be an indicator for tumor aggressiveness and a parameter to be taken into consideration in tumor therapy. To analyze COX-2 expression and PGE2 synthesis, we have developed a novel flow cytometric assay, which allows the determination of COX-2 expression and PGE2 synthesis simultaneously. We have studied the in vitro influence of the TNF-alpha cytokine on COX-2 expression and PGE2 synthesis in human melanoma cells (A375). In addition, we have performed a clinical study on tumor tissue and blood of patients with primary malignant melanoma. We found that in vitro stimulation with TNF-alpha results in a rapid and a sustained COX-2 expression and in a corresponding increased PGE2 synthesis. In vivo, we observed a continuous increase of COX-2 expression and PGE2 synthesis in tumor tissue of patients with primary malignant melanoma. We were able to demonstrate that the levels of PGE2 in blood, measured by our assay, were similar to those determined by the enzyme-linked immunosorbent assay (ELISA). This study shows that COX-2 expression and PGE2 synthesis can be determined simultaneously by flow be359ba680
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