Premium
Zinc Deficiency Induces Apoptosis but Zinc Induces Necrosis in Rat Hepatoma Cells
Author(s) -
Sankavaram Kavitha,
Chong Leelyn,
Bruno Richard S,
Freake Hedley C
Publication year - 2010
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.24.1_supplement.718.3
Subject(s) - apoptosis , programmed cell death , necrosis , zinc , annexin , zinc deficiency (plant disorder) , reactive oxygen species , flow cytometry , microbiology and biotechnology , biology , tumor necrosis factor alpha , fragmentation (computing) , viability assay , dna fragmentation , chemistry , biochemistry , immunology , ecology , genetics , organic chemistry
Altered concentrations of intracellular zinc influence cellular homeostasis and are believed to modulate apoptosis. We reported earlier that both zinc supplementation and DTPA (diethylenetriaminepentaacetic acid) induced zinc deficiency generate reactive oxygen species (ROS) and decrease cell viability in rat hepatoma (H4IIE) cells. In this study, we examined the effects of altered zinc availability on mode of cell death, signaling pathways and associated morphological changes in these cells. Fluorescence microscopy of cells with Hoechst 33342 and PI indicated that DTPA induced apoptosis but zinc induced both necrosis and apoptosis. Cells were also stained with both PI and FTTC‐labeled annexin V and then analyzed by flow cytometry. The results confirmed that DTPA induced apoptosis (20%) whereas zinc induced both necrosis (50%) and apoptosis (10%). Zinc induced necrosis prior to apoptosis and at a greater rate. Features of apoptosis, such as chromatin condensation, nuclear fragmentation, elevated levels of p53, p21, and Fas/Fas ligand mRNA accompanied DTPA and zinc induced cell death. Decreased expression of Bcl2 and elevated expression of Bax were observed with DTPA but not with zinc treatment. These results indicate that zinc deficiency leads to mitochondrial dependent apoptosis, modulating ROS and apoptosis proteins in H4IIE cells. However, elevated zinc leads predominantly to cell necrosis. Grant Funding Source : UConn Research Foundation