ABSTRACT
Volatile organic compounds (VOCs) such as benzene and toluene, and low molecular weight carbonyls like formaldehyde belong to the main air pollutants found in indoor environments. They are suspected to induce acute and chronic adverse health effects like asthma, allergic and cardiovascular diseases, and strongly affect well-being. Our aim was to further develop and optimize an in vitro method to study the exposure of epithelial tumour lung cells (A549) by using a commercial exposure chamber (CULTEX) to assess the biological effects of VOCs and carbonyl compounds at low concentration levels. Exposing the cells to toluene, benzene and formaldehyde at mixing ratios varying from 0.1 to 0.6ppmv in air resulted in reproducible direct effects with the induction of an inflammatory response and a modification of the glutathione redox status.
Subject(s)
Cell Culture Techniques/methods , Respiratory Mucosa/drug effects , Volatile Organic Compounds/toxicity , Air Pollution, Indoor/adverse effects , Cell Line, Tumor , Cell Membrane/drug effects , Cell Membrane/pathology , Glutathione/metabolism , Humans , Inflammation/chemically induced , Interleukin-8/metabolism , L-Lactate Dehydrogenase/metabolism , Respiratory Mucosa/metabolismABSTRACT
Human epithelial lung cells (A549) were exposed to toluene and benzene in the air as individual compounds and mixtures at concentrations of about 0.25ppmv in a specifically adapted fumigation device. Possible early toxicological effects at cellular level have been determined by lactate dehydrogenase (LDH), glutathione redox status (GSH) and comet assay. An hour of exposure to 0.25ppmv of toluene in the air induced DNA damages which were repaired within 24h after the treatment. No DNA damage was detected by applying a similar concentration of benzene, but there was a decrease in the glutathione ratio. Exposure to a mixture of toluene and benzene in air led to an increase in the cytotoxic effect and DNA damage without any further repair, but did not induce any changes in the glutathione redox status.
Subject(s)
Air Pollution, Indoor/adverse effects , Benzene/toxicity , Epithelial Cells/drug effects , Lung/drug effects , Toluene/toxicity , Benzene/chemistry , Cell Line , Cell Survival/drug effects , Comet Assay , DNA Damage , DNA Repair , Drug Synergism , Epithelial Cells/metabolism , Epithelial Cells/pathology , Glutathione/metabolism , Humans , Lung/cytology , Oxidation-Reduction , Toluene/chemistryABSTRACT
A new murine cell line, named GFPneu, was established from a mammary adenocarcinoma arising in double transgenic MMTVneu x CMV-GFP mice. Breast tumours develop in 100% of females after 2 months latency, as a result of the over-expression of the activated rat neu oncogene in the mammary glands. All tissues, and in particular the breast tumours, express the GFP protein. This cell line was tumorigenic when inoculated into nude mice and the derived tumours showed the same histological features as the primaries from which they were isolated. Their histopathology reproduces many characteristics of human breast adenocarcinomas, in particular their ability to metastasize. The GFP marker allows us to visualize the presence of lung metastases in fresh tissues immediately, to confirm the histopathology. From a lung metastatic fluorescent nodule, we derived a further cell line, named MTP-GFP, which we also characterized. These two cell lines could be useful to study the role played by the neu oncogene in the maintenance of the transformed phenotype, in the metastatic process, to test novel therapeutic strategies to inhibit primary tumour growth and to observe the generation of distant metastases.
Subject(s)
Adenocarcinoma/genetics , Cell Line, Tumor , Genes, erbB-2/genetics , Green Fluorescent Proteins/genetics , Mammary Neoplasms, Animal/genetics , Adenocarcinoma/secondary , Animals , Female , Gene Expression Regulation, Neoplastic , Lung Neoplasms/genetics , Lung Neoplasms/secondary , Mammary Neoplasms, Animal/pathology , Mice , Mice, Nude , Mice, Transgenic , TelomereABSTRACT
TNFR1/Fas engagement results in the cleavage of cytosolic Bid to truncated Bid (tBid), which translocates to mitochondria. We demonstrate that recombinant tBid induces in vitro immediate destabilization of the mitochondrial bioenergetic homeostasis. These alterations result in mild uncoupling of mitochondrial state-4 respiration, associated with an inhibition the adenosine diphosphate (ADP)-stimulated respiration and phosphorylation rate. tBid disruption of mitochondrial homeostasis was inhibited in mitochondria overexpressing Bcl-2 and Bcl-XL. The inhibition of state-3 respiration is mediated by the reorganization of cardiolipin within the mitochondrial membranes, which indirectly affects the activity of the ADP/ATP translocator. Cardiolipin-deficient yeast mitochondria did not exhibit any respiratory inhibition by tBid, proving the absolute requirement for cardiolipin for tBid binding and activity. In contrast, the wild-type yeast mitochondria underwent a similar inhibition of ADP-stimulated respiration associated with reduced ATP synthesis. These events suggest that mitochondrial lipids rather than proteins are the key determinants of tBid-induced destabilization of mitochondrial bioenergetics.
