The expression of aromatase is tissue-specifically regulated through the alternative use of multiple exons 1 and promoters. We analysed expression levels of aromatase mRNA, preferential utilization of multiple exon 1 of the human aromatase gene, and transcriptional regulation of their multiple promoters in breast cancer tissues by newly developed fluorometric methods. The expression levels of aromatase mRNA in breast cancer tissues were significantly higher than those in regions distal to tumours or in non-malignant breast tissues. Aromatase mRNA in these non-malignant tissues was transcribed from skin fibroblast/fetal liver-specific exon 1 (exon 1b) of the aromatase gene. However, in half the cases of breast cancer patients, the utilization of multiple exons 1 in the aromatase mRNA changed from exon 1b to ovary-specific exon 1 (exon 1c) in their breast tissues. Aromatase mRNA in HepG2 cells as well as in non-malignant breast tissues was also transcribed from exon 1b. Then, the promoter region responsible for the exon 1b-specific utilization in HepG2 cells was examined by fluorometric promoter assay using a new reporter containing four major alternative exons 1 and promoters. The results suggested that transcriptional elements determining preferential utilization of exon 1b in the cells was located on the promoter region of exon 1b from -255 to -1145. To investigate further the cause of the elevation of aromatase mRNA and the switching from exon 1b to exon 1c in the transcription of the aromatase gene, the effects of various factors on the expression levels and preference of alternative exons 1 were examined in cultured adipose stromal cells from breast tissues. Aromatase mRNA was transcribed from exon 1b in the stromal cells, cultured in the presence of calf serum. However, removal of the serum or the addition of forskolin or phorbol ester (TPA) induced a rapid elevation of aromatase mRNA and the switching of aromatase transcripts to exon 1c in the cells, whereas TGFbeta almost abolished the expression of aromatase mRNA. Because co-culture of cancer cells such as MCF-7 increased aromatase mRNA of the cells cultured in the serum-containing medium, it is possible that cancer cells secret stimulatory factors acting like forskolin or TPA, or consume serum inhibitory factors acting like TGFbeta, consequently causing levels of aromatase mRNA to increase.