Phorbol diester 12-O-tetradecanoylphorbol 13-acetate (TPA) up-regulates the expression of estrogen receptors in human THP-1 leukemia cells.
In the present work, we have inspected expression of estrogen receptors (ER) and their regulation by the phorbol diester 12-O-tetradecanoylphorbol 13-acetate (TPA) in a leukemic cell line, the THP-1 cells, using multiple experimental approaches. Firstly, ligand binding assay (LBA) revealed that control (unstimulated) THP-1 cells express type I (high affinity, limited capacity) ER in the nuclear fraction only, whilst treatment of cells with TPA resulted in the appearance of type I ER in the soluble fraction as well, with the 50 ng/ml dose and the 48 h incubation time being the most effective experimental condition. A concomitant increase of type II ER was also seen in both soluble and nuclear cell fractions. Unstimulated THP-1 cells were found to be ER negative by immunocytochemistry; conversely, cells exposed to 50 ng/ml TPA for 48 h stained positively for ER, with the majority of cells having a strong nuclear staining. Scrutiny of ER mRNA expression using reverse transcriptase-polymerase chain reaction showed the presence of a wild type ER transcript in both control and TPA-treated THP-1 cells, though levels of ER mRNA were found to be comparatively higher in the latter. This combined evidence would imply that the TPA-induced differentiation of THP-1 cells is accompanied by the rise of high affinity (type I) ER, suggesting that estrogens may play a role in the regulation of macrophage activity during the inflammatory and/or the immune response.
Cutolo
M
M
Division of Rheumatology, Department of Internal Medicine, University of Genoa, Italy.
Carruba
G
G
Villaggio
B
B
Coviello
D A
DA
Dayer
J M
JM
Campisi
I
I
Miele
M
M
Stefano
R
R
Castagnetta
L A
LA
D002454:Cell Differentiation; D002467:Cell Nucleus / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007150:Immunohistochemistry; D007249:Inflammation; D007700:Kinetics; D007938:Leukemia / Q000378:metabolism*; D008024:Ligands; D012333:RNA, Messenger / Q000378:metabolism; D011960:Receptors, Estrogen / Q000096:biosynthesis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D013997:Time Factors; D014407:Tumor Cells, Cultured; D015854:Up-Regulation
D008024:Ligands; D012333:RNA, Messenger; D011960:Receptors, Estrogen; D013755:Tetradecanoylphorbol Acetate