Mechanism of apoptosis and determination of cellular fate in chromium(VI)-exposed populations of telomerase-immortalized human fibroblasts.
The cellular responses to carcinogen exposure influence cellular fate, which in turn modulates the neoplastic response. Certain hexavalent chromium [Cr(VI)] compounds are implicated as occupational respiratory carcinogens at doses that are both genotoxic and cytotoxic. We examined the mechanism of Cr(VI)-induced apoptosis in normal human fibroblasts (BJ) immortalized by human telomerase gene transfection (BJ-hTERT), and we assessed the spectrum of cumulative cellular fates [(a) regaining of replicative potential; (b) terminal growth arrest; or (c) apoptosis] for a narrow range of increasingly genotoxic doses of Cr(VI). Exposure of BJ-hTERT cells to Cr(VI) resulted in a dose-dependent increase in apoptosis that involved mitochondrial disruption as evidenced by mitochondrial membrane depolarization and cytochrome c release. The initial response to Cr(VI) exposure was inhibition of cell cycle progression. At the lowest dose tested (1 microM; 32% clonogenic survival), the cell cycle inhibition led to terminal growth arrest but no apoptosis. The fraction of terminally growth arrested cells increased as the dose was increased to 3 microM but then decreased at 4, 5, and 6 microM as apoptosis became the predominant cell fate. Our results suggest that cell populations exposed to Cr(VI) have a different spectrum of responses, depending on the extent of DNA damage, and that the regaining of replicative potential after relatively higher genotoxic exposures may be attributable to either escape from, or resistance to, terminal growth arrest or apoptosis.
Pritchard
D E
DE
Department of Pharmacology, The George Washington University Medical Center, Washington, DC 20037, USA.
Ceryak
S
S
Ha
L
L
Fornsaglio
J L
JL
Hartman
S K
SK
O'Brien
T J
TJ
Patierno
S R
SR
D017209:Apoptosis; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002453:Cell Cycle; D002455:Cell Division / Q000187:drug effects; D019070:Cell Lineage; D002478:Cells, Cultured; D002840:Chromates / Q000494:pharmacology*; D004268:DNA-Binding Proteins; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000187:drug effects* / Q000473:pathology*; D006801:Humans; D007150:Immunohistochemistry; D007425:Intracellular Membranes; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000378:metabolism; D010641:Phenotype; D017670:Sodium Compounds / Q000494:pharmacology*; D013312:Stress, Physiological; D019098:Telomerase / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection
D002840:Chromates; D004268:DNA-Binding Proteins; D017670:Sodium Compounds; C028982:sodium chromate(VI); D019098:Telomerase; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases