Effects of Conditioned Medium from Different Cultured Cell Types on Aromatase Expression in Adipose Stromal Cells

Abstract

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 17β-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 underfined, heat-labile and proteinaceous factors are present in CM that stimulate P450arom expression in a fashion similar to that of serum.

Keywords

Aromatase adipose conditioned medium breast cancer

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