Effects of conditioned medium from different cultured cell types on aromatase expression in adipose stromal cells.
OBJECTIVE
To determine whether serum-free (SF) conditioned media (CM) from several human breast cancer cell lines and primary stromal cell cultures contain factor(s) that mimic the marked stimulatory effects of serum on aromatase activity and aromatase P450 (P450arom) gene expression in adipose stromal cells in culture (ASC) in the presence of dexamethasone (DEX).


METHODS
Adipose stromal cells, harvested from fresh adipose specimens, were grown to confluence, switched to SF media, and then incubated in the presence or absence of DEX with CM from T47-D breast cancer cells, pre-treated with or without 17 beta-estradiol (E2), and with CM from stromal cell cultures. Aromatase activity of the ASC was determined by the [3H] water release assay. Total RNA was isolated, and reverse transcription-polymerase chain reaction was performed to determine the expression of various 5'-termini.


RESULTS
T47-D CM stimulated aromatase activity in a concentration-dependent manner, similar to that of serum, in ASC incubated with DEX. Estrogen potentiated this in a dose-dependent fashion. The ASC CM and endometrial stromal cell CM also markedly induced aromatase activity in ASC. Heat inactivation destroyed the stimulating ability of CM. The majority of P450arom 5'-termini expressed by ASC incubated with CM plus DEX contained the promoter I.4-specific sequence.


CONCLUSIONS
Conditioned media from several breast cancer cell lines and primary stromal cell cultures can mimic the effects of serum in the presence of DEX to stimulate aromatase activity in ASC. These results suggest that undefined, heat-labile and proteinaceous factors are present in CM that stimulate P450arom expression in a fashion similar to that of serum.
Nichols
J E
JE
Cecil H. and Ida Green Center for Reproductive Biology Sciences, University of Texas Southwestern Medical Center, Dallas 75235-9051, USA.
Bulun
S E
SE
Simpson
E R
ER
D000273:Adipose Tissue / Q000166:cytology* / Q000201:enzymology*; D001141:Aromatase / Q000096:biosynthesis*; D001943:Breast Neoplasms; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D017931:DNA Primers; D003907:Dexamethasone / Q000494:pharmacology; D004958:Estradiol / Q000031:analogs & derivatives / Q000494:pharmacology; D004965:Estrogen Antagonists / Q000494:pharmacology; D005260:Female; D000077267:Fulvestrant; D006801:Humans; D016133:Polymerase Chain Reaction; D017154:Stromal Cells / Q000201:enzymology; D014407:Tumor Cells, Cultured
D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D017931:DNA Primers; D004965:Estrogen Antagonists; D000077267:Fulvestrant; D004958:Estradiol; D003907:Dexamethasone; D001141:Aromatase