Submitted to: In Vitro Biology Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 2/6/2008
Publication Date: 6/1/2008
Citation: Stanley, D.W., Goodman, C.L., Song, Q., An, S., Mcintosh, A.H. 2008. Prostaglandin A2 significantly alters gene expression in an established insect cell line (BCIRL-HzAM1) [abstract]. In Vitro Biology Meeting. p. S43
Technical Abstract: In previous work to determine the biochemical mechanisms of prostaglandin (PG) action in insect cells, we found that PGA1 and PGE1 influenced the expression of genes encoding proteins important for a variety of cellular functions. In the present study, we exposed the same cell line, BCIRL-HzAM1, to three PGs of the 2-series to determine if they would have similar effects. Cells were incubated with 15 uM PGA2 (which interacts with perinuclear receptors), PGE2, or PGF2a'(both of which act via G protein coupled receptors) for 12 or 24 hr. Differences in protein patterns were determined quantitatively using 2D electrophoresis. Dramatic changes in protein expression were noted in cells exposed to PGA2 for 12 hr, with significant changes in over 70 proteins. Modifications in protein expression were also noted for the PGE2 or PGF2a treated cells, but not nearly to the same extent. Identities for some of the proteins of interest had been previously elucidated in our earlier study using MS/MS (MALDI TOF/TOF). Therefore, we can at this point clearly state that PGA2 significantly alters the expression of proteins involved in numerous cellular functions, including cell protection (e.g., catalase and various superoxide dismutases), cell movement/division (e.g., actin-depolymerization factor), protein action (e.g., heat shock cognate), lipid metabolism (e.g., acyl-coenzyme A dehydrogenase), signal transduction (e.g., 14-3-3 protein), and metabolism (e.g., arginine kinase and enolase). We are identifying more proteins via MS/MS to expand our knowledge of those that are up- or down-regulated by PGs, as well as confirming the changes in levels of specific proteins using qPCR. Our results continue to give strong support for our hypothesis that PGs influence gene expression in insect cells.