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Overexpression of basic fibroblast growth factor (FGF–2) downregulates Bcl–2 and promotes apoptosis in MCF–7 human breast cancer cells

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Abstract

Basic fibroblast growth factor (bFGF, FGF–2), a classical transforming factor, mitogen, and survival factor in multiple cell types, and has a paradoxic role in mammary epithelial cell transformation and proliferation. We have also demonstrated that recombinant FGF–2 uncharacteristically promotes cell death in MCF–7 human breast cancer cells. In this study, we investigated the effects of FGF–2 overexpression on survival in the same MCF–7 cells. In eight breast cancer cell lines and two nontransformed mammary epithelial cell lines, we demonstrated that high levels of Bcl–2 are only expressed in cells with undetectable levels of FGF–2 on western blot. In retrovirally transduced MCF–7 cells expressing both cytoplasm– and nucleus–localizing FGF–2 species and ones expressing only cytoplasm–localizing FGF–2 species, Bcl–2 levels were strongly decreased at both the mRNA and protein levels. Immunoprecipitation of Bax demonstrated a decreased association of Bax with Bcl–2 in these cells. Levels of Bax did not correlate with expression of FGF–2 in the 10 cell lines or in MCF–7 cells. The clonogenic potential of MCF–7 cells in tissue culture was decreased by the expression of FGF–2 and was additively suppressed by the chemotherapeutic agents etoposide and 5–fluorouracil in a dose and time dependent manner. MCF–7 cells overexpressing FGF–2 had a greater rate of programmed cell death at baseline and in response to etoposide and 5–fluorouracil in a TUNEL assay by immunofluorescent microphotography and by flow cytometric quantitation. The pro–apoptotic effect of FGF–2 overexpression on the chemosensitivity of these cells was confirmed by quantitative morphologic determination. These data demonstrate that the expression of FGF–2 downregulates Bcl–2 and promotes programmed cell death in MCF–7 human breast cancer cells.

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Authors and Affiliations

  1. Division of Oncology, Department of Medicine, USA

    Paul Maloof, Qin Wang, Huisheng Wang & Robert Wieder

  2. Department of Pediatrics, UMDNJ–New Jersey Medical School, Newark, NJ, USA

    Dana Stein & Thomas N. Denny

  3. Department of Radiation Oncology, Memorial Sloan–Kettering Cancer Center, New York, NY, USA

    Joachim Yahalom

  4. Felsenstein Medical Research Center, Rabin Medical Center, Beilinson Campus, Petach–Tikva, Israel

    Eyal Fenig

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  1. Paul Maloof
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  2. Qin Wang
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  3. Huisheng Wang
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  4. Dana Stein
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  5. Thomas N. Denny
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  6. Joachim Yahalom
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  7. Eyal Fenig
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  8. Robert Wieder
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Maloof, P., Wang, Q., Wang, H. et al. Overexpression of basic fibroblast growth factor (FGF–2) downregulates Bcl–2 and promotes apoptosis in MCF–7 human breast cancer cells. Breast Cancer Res Treat 56, 151–165 (1999). https://doi.org/10.1023/A:1006258510381

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