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Fibroblast Growth Factor‐2 inhibits Bleomycin‐induced DNA Damage in Murine Lung Endothelial Cells.
Author(s) -
Stevens Rachel L.,
Okoreeh Andre,
Hoyt Dale G.
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a673-c
Subject(s) - bleomycin , cancer research , chemistry , fibroblast growth factor , dna damage , microbiology and biotechnology , lung , fibroblast , dna , biology , medicine , biochemistry , in vitro , chemotherapy , receptor
Fibroblast Growth Factor‐2 (FGF2) protects mice from radiation‐ and LPS‐induced lung injury. Here we investigated the effect of FGF2 on DNA damage caused by bleomycin (BLM) in mouse lung endothelial cells (MLEC). MLEC were treated with 0.1‐10 ng FGF2/ml for 1 to 4h, and then with 100 or 500 ug BLM/ml for 15‐45 min. Cells were either fixed with formaldehyde and permeabilized in 70% ethanol, or fixed with 95% ethanol/1% acetic acid and stored in PBS. In formaldehyde‐fixed cells, DNA strand breaks containing 3′OH were labeled with fluorescein‐12‐dUTP by nick translation with DNA polymerase. Ethanol/acetic acid‐fixed cells were immunostained with antibody to the phosphorylated histone (H) 2A variant, H2Ax (phospho‐H2Ax), which accumulates at sites of double strand breakage, and a Cy3‐secondary antibody. Cells were finally stained with H33342 to mark DNA. Fluorescence microscopic image intensities of fluorescein for 3′OH labeling, or Cy3 for phospho‐H2Ax, were analyzed. FGF2 inhibited both measures of strand breakage. The results suggest that FGF2 can protect lung EC from BLM‐induced single and/or double strand DNA breakage. Support: NIH HL68054.