Regulation of a promoter of the fibroblast growth factor 1 gene in prostate and breast cancer cells.
FGF-1 mRNA is expressed in the prostate cancer cell lines LNCaP and PC-3 and in the breast carcinoma cell line MDA-MB-231. Levels of FGF-1 mRNA have been shown to be up-regulated by serum, phorbol esters, and combinations of growth factors. It was shown that the major FGF-1 mRNA species expressed following serum stimulation in MDA-MB-231 cells is FGF-1.C. To better understand the potential role of FGF-1 in human prostate and breast cancer, we began an analysis of the cis- and trans-acting elements of one of its promoters required for the serum, PMA, and androgen regulation in breast and prostate cancer cell lines. We show that FGF-1.C steady-state mRNA levels are increased following serum or PMA stimulation of PC-3 cells. Further, we determine the FGF-1.C transcription start site in PC-3 cells. By sequence analysis, we show that consensus AP1, AP2, and Sp1 sites and ARE- and CRE-near consensus elements are present in the immediate 5' region of the FGF-1.C transcription start site. Gel-shift assays show that oligonucleotides containing FGF-1.C AP1, AP2, or Spl sequences form specific DNA-protein complexes with nuclear extracts from PC-3 cells. To determine if these or other cis-acting sequences are responsible for the serum, androgen, or growth factor regulation of FGF-1 expression, fragments of the FGF-1.C promoter region were cloned upstream of the luciferase reporter gene. We show that FGF-1 synergizes with androgen to enhance FGF-1.C transcription in LNCaP cells. We further show that the DNA fragment containing sequence up to 1614 nucleotides upstream of the FGF-1.C transcription start site is sufficient for stimulating promoter activity following serum treatment of MDA-MB-231 cells. Thus, FGF-1.C promoter contains sequences that are important for androgen or serum stimulation in prostate and breast cancer cells.
Department of Internal Medicine and Comprehensive Cancer Center, The Ohio State University, Columbus 43210, USA.
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Payson
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Chotani
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Chiu
D000728:Androgens / Q000494:pharmacology; D001483:Base Sequence; D001769:Blood; D001943:Breast Neoplasms / Q000235:genetics*; D002460:Cell Line; D016384:Consensus Sequence; D018076:DNA, Complementary / Q000737:chemistry; D016220:Fibroblast Growth Factor 1 / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011471:Prostatic Neoplasms / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D017421:Sequence Analysis; D013755:Tetradecanoylphorbol Acetate; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured
D000728:Androgens; D018076:DNA, Complementary; D012333:RNA, Messenger; D016220:Fibroblast Growth Factor 1; D013755:Tetradecanoylphorbol Acetate