Effects of phorbol esters on an interleukin-3-dependent cell line

Abstract

FDC-P1 is an interleukin-3 (IL-3)-dependent cell line that ceases to proliferate in the absence of IL-3. We have isolated variant cell lines from FDC-P1 that grow in response to phorbol myristate acetate (PMA). These variant cell lines (FD/PMA) have maintained their PMA-dependency for over 1 year. Lymphokine gene expression, which would support growth, was not detected in FD/PMA lines. FD/PMA lines had a different cell surface phenotype than the parental line. Mac-1, Mac-2, and Mac-3 were readily detected on the cell surface of FD/PMA lines; however, these antigens were not detected on FDC-P1. Because protein kinase C (PKC) activation may mediate PMA effects, we examined this kinase. PKC activity quantitated by 32P-incorporation into histone was increased in FDC-P1 as compared with FD/PMA cultured in IL-3. Moreover, PKC activity was undetectable in FD/PMA lines cultured in PMA. Using Western blotting, immunoreactive PKC was readily detected in cytosolic and solubilized particulate fractions of FDC-P1 cells, not but in FD/PMA cell extracts. Comparisons between the parental and FD/PMA lines should provide insight into IL-3- and PMA-mediated signal transduction.