Stress response of mammalian cells incubated with landfill leachate.
Environmental contamination from landfill leachate has been linked to disturbances in human health. Often, however, only global parameters, such as dissolved organic content, chemical oxygen demand, and 5-d biological oxygen demand, are used to evaluate wastewater quality. In the present study, we determined leachate cytotoxicity and stress response of leachate-treated mammalian cells using in vitro bioassays and other molecular techniques. The modified E-screen assay using human breast cancer MCF-7 cells was used to determine the estrogenic potential and/or cytotoxicity of water samples from two solid-waste landfills in Tunisia. The cytotoxicity mechanism of the leachate was determined by DNA fragmentation and lactate dehydrogenase assays. The stress response of heat shock protein (HSP) 47-positive Chinese hamster ovary cells treated with leachate also was determined. Proteomics analyses and real-time reverse transcription-polymerase chain reaction were used to determine and confirm the enhanced expressions of certain stress-related proteins. Results showed that the leachate samples generally did not have estrogenic activity. Instead, they were cytotoxic toward MCF-7 cells, and the cytotoxicity was by necrosis during the early stages of incubation. Leachate also enhanced the expression of HSP and various stress-related proteins, such as heterogeneous nuclear ribonucleoprotein E1, phosphoglycerate mutase, and nuclear matrix protein 200, in MCF-7 cells. These can be considered as survival mechanisms against leachate-induced cytotoxicity.
Talorete
Terence
T
Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba City, Ibaraki, Japan.
Limam
Atef
A
Kawano
Mitsuko
M
Ben Rejeb Jenhani
Amel
A
Ghrabi
Ahmed
A
Isoda
Hiroko
H
D000818:Animals; D001483:Base Sequence; D045744:Cell Line, Tumor; D053938:DNA Fragmentation; D017931:DNA Primers; D050958:HSP47 Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D040901:Proteomics; D012037:Refuse Disposal; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014874:Water Pollutants, Chemical / Q000633:toxicity*
D017931:DNA Primers; D050958:HSP47 Heat-Shock Proteins; D014874:Water Pollutants, Chemical; D007770:L-Lactate Dehydrogenase