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Antioxidant function of polyamines in human colon cancer cells demonstrated by fluorescence
Author(s) -
Thornton Jonathan M,
Osborne David L
Publication year - 2012
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.26.1_supplement.692.15
Subject(s) - putrescine , polyamine , reactive oxygen species , antioxidant , biochemistry , fluorescence , cancer cell , spermidine , biology , chemistry , microbiology and biotechnology , cancer , enzyme , physics , genetics , quantum mechanics
Polyamines are known to be essential to the growth of eukaryotic cells. Studies have shown that polyamines may prevent oxygen mediated damage. We postulate that a possible function for polyamines in mature enterocytes may relate to an antioxidant function. The focus of this study was to visualize reactive oxygen species under various treatment conditions and determine if alterations in polyamine content may influence ROS mediated fluorescence. Studies used human colon cancer cells (HCT116) grown in culture. Cells were grown using RPMI1640 medium + 10% FCS. Media was supplemented with DFMO, putrescine or DFMO + putrescine. Cells were grown for 24 hours before fluorescent analysis was performed. ROS activity was visualized by using the Image‐iT® fluorescent dye… Cells were loaded with dye and then exposed to 6 mM H 2 O 2 . In the cells treated with RPMI1640 + 10% FCS, a baseline fluorescence was observed. Addition of DFMO resulted in a marked increase in the observed fluorescence compared to the control. Addition of putrescine to the DFMO media demonstrated that putrescine could reverse the affects of DFMO treatment. These results indicate that when polyamines are present, human colon cancer cells have a reduced observable load of ROS while the ROS load is increased in the conditions of reduced polyamine levels. This supports the theory that polyamines are functioning as antioxidants in gastrointestinal cells.