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Is available on suppression of these genes by particles. We show herein an unexpected and pronounced reduction of CYP expression in various cell types. The reduction of CYP protein may lead to disruption of various cellular functions: pathophysiological response tostress signals (toxic substances or inflammation), an adaptive homeostatic response (controlled generation of reactive oxygen species) or a part of tightly regulated physiological pathway (production of bile acid) . Inflammatory mediators can be responsible for a reduction of CYP mRNA . Ke et al. reported that Staurosporine one mechanistic explanation for LPS and cytokine-mediated CYP reduction may be an interaction with NFB . The reduction of CYP1B1 after ultrafine P90 treatment could be a form of protective mechanism concerning oxidative stress. Reactive oxygen species (ROS) generated by CYP-catalyzed metabolisms can cause oxidative stress in cells . Reduced CYP1B1 may prevent further DNA damage being caused by ROS. Furthermore, decreased availability of CYP1B1 enzyme leads to a lesser activation of PAH to reactive metabolites which otherwise could cause DNA damage and cancer development . On the other hand CYP1B1 and CYP1A1 are also important for detoxifying various xenobiotics. A reduced PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/25751659 expression of these cytochromes in monocytes/macrophages and lung epithelial cells induced by inhaled carbon particles may lead to increased damage by xenobiotics because of the lack of its detoxifying character. Further investigations will be required to elucidate by which mechanism the particles affect the CYP1B1 expression.MethodsCell lines and culture medium The epithelial cell lines A549  and Calu-3  were cultured in Dulbecco's minimal essential medium NUT mix F12 (21331-020, Invitrogen, Karlsruhe, Germany), containing 10 FCS (S0115, Biochrom, Berlin, Germany), L-glutamine 2 mM (15140-114, Invitrogen, Karlsruhe, Germany), and penicillin 200 U/ml/streptomycin 200 g/ml (15140-114, Invitrogen, Karlsruhe, Germany).Primary human cells were cultured in RPMI 1640 medium (F1415, Biochrom, Berlin, Germany), supplemented with OPI (oxaloacetate, pyruvate, insulin; O5003, Sigma, Taufkirchen, Germany), L-glutamine 2 mM (15140-114, Invitrogen, Karlsruhe, Germany), penicillin 200 U/ml/streptomycin 200 g/ml (15140-114, Invitrogen, Karlsruhe, Germany), and 1?nonessential amino acids (11140-35, Invitrogen, Karlsruhe, Germany). Before addition of 10 FCS (S0115, Biochrom, Berlin, Germany), the supplemented medium was ultrafiltered through a Gambro 2000 column (Gambro, Hechingen, Germany) in order to remove any inadvertent LPS contamination.Donors of primary human cells Primary human cells were obtained from healthy human volunteers (non-smoker and smoker), from patients with COPD, or from patients with other lung diseases (TablePage 9 of(page number not for citation purposes)Particle and Fibre Toxicology 2009, 6:http://www.particleandfibretoxicology.com/content/6/1/Table 1: Characteristics of patients and volunteers from which primary human cells were obtained.number normal healthy healthy smokers COPD patients others (brush biopsies) 23 6 12sexagedisease and stage n.a. n.a. 4?II, 3?III, 5?IV (Gold) adenocarcinoma, MALT-lymphoma, UIP, sarcoidosis (Gold II), COPD (Gold III)*12?m, 11?f 42.8 ?15.4 4?m, 2?f 54.5 ?10.6 10?m, 2?f 63.9 ?10.5 5?m, 1?f, 1?anonymized* 65.8 ?11.7*(*: samples from one patient who underwent brush biopsy was anonymized; f: female; m: male; MALT: mucosa-associated lymph.
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