Molecular analysis of aberrant expression of aromatase in breast cancer tissues.

Aromatase mRNA in non-malignant breast tissues was mainly transcribed form skin fibroblast/fetal liver-specific exon 1 (exon 1b) of the aromatase gene. However, in half the cases of breast cancers, switching of the alternative exons 1 from exon 1b to ovary-specific exon 1 (exon 1c) was observed, and expression levels of aromatase mRNA in breast cancer tissues were significantly higher than those in the distal regions to tumors or in non-malignant breast tissues. Co-culture or addition of the conditioned medium of breast cancer cells, MCF-7 caused increase of aromatase mRNA in cultured adipose stromal cells from breast tissues together with switching from exon 1b to exon 1c. Removal of fetal calf serum (FCS) from the culture medium or addition of forskolin or phorbol ester (TPA) also induced rapid elevation of aromatase mRNA and switching to exon 1c, whereas TGFbeta almost abolished the expression, suggesting that cancer cells might secret forskolin- or TPA-like stimulatory factors, or consume TGFbeta-like inhibitory factors in serum for expression of aromatase mRNA. The promoter region responsible for transcription from exon 1b and the switching was investigated using a newly developed reporter carrying 4 major alternative exons 1 and promoters. Transcriptional elements responsible for preferential utilization of exon 1b were identified on the promoter region between -300 and -400 of exon 1b. Gel shift assay showed a unique site for a FCS-dependent DNA binding factor just downstream of GRE (glucocorticoid responsive element) in this promoter region.