ABSTRACT
Drugs that target novel surfaces on the androgen receptor (AR) and/or novel AR regulatory mechanisms are promising alternatives for the treatment of castrate-resistant prostate cancer. The 52 kDa FK506 binding protein (FKBP52) is an important positive regulator of AR in cellular and whole animal models and represents an attractive target for the treatment of prostate cancer. We used a modified receptor-mediated reporter assay in yeast to screen a diversified natural compound library for inhibitors of FKBP52-enhanced AR function. The lead compound, termed MJC13, inhibits AR function by preventing hormone-dependent dissociation of the Hsp90-FKBP52-AR complex, which results in less hormone-bound receptor in the nucleus. Assays in early and late stage human prostate cancer cells demonstrated that MJC13 inhibits AR-dependent gene expression and androgen-stimulated prostate cancer cell proliferation.
Subject(s)
Gene Expression Regulation/drug effects , HSP90 Heat-Shock Proteins/metabolism , Models, Molecular , Multiprotein Complexes/metabolism , Prostatic Neoplasms/metabolism , Receptors, Androgen/metabolism , Tacrolimus Binding Proteins/antagonists & inhibitors , Animals , Cell Line, Tumor , Cell Proliferation/drug effects , Drug Discovery , Enzyme-Linked Immunosorbent Assay , Fluorescence , Humans , Immunoblotting , Immunoprecipitation , Male , Mice , Molecular Dynamics Simulation , Molecular Structure , Receptors, Androgen/chemistry , Tacrolimus Binding Proteins/metabolism , Yeasts , beta-GalactosidaseABSTRACT
Aerosol particulates collected on filters from ballistic penetration and erosion events for W-Ni-Co and W-Ni-Fe kinetic energy rod projectiles penetrating steel target plates were observed to be highly cytotoxic to human epithelial A549 lung cells in culture after 48 hours of exposure. The aerosol consisted of micron-sized Fe particulates and nanoparticulate aggregates consisting of W, Ni or W, Co, and some Fe, characterized by scanning electron microscopy and transmission electron microscopy, and using energy-dispersive (X-ray) spectrometry for elemental analysis and mapping. Cytotoxic assays of manufactured micron-sized and nanosized metal particulates of W, Ni, Fe, and Co demonstrated that, consistent with many studies in the literature, only the nanoparticulate elements demonstrated measurable cytotoxicity. These results suggest the potential for very severe, short-term, human toxicity, in particular to the respiratory system on inhaling ballistic aerosols.
Subject(s)
Metal Nanoparticles/chemistry , Metal Nanoparticles/toxicity , Respiratory System/drug effects , Aerosols , Cell Line , Cell Survival/drug effects , Cobalt/toxicity , Forensic Ballistics , Humans , Iron/toxicity , Metal Nanoparticles/administration & dosage , Metal Nanoparticles/ultrastructure , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , Nanomedicine , Nickel/toxicity , Particle Size , Tungsten/toxicityABSTRACT
The nature and constituents of ballistic aerosol created by kinetic energy penetrator rods of tungsten heavy alloys (W-Fe-Ni and W-Fe-Co) perforating steel target plates was characterized by scanning and transmission electron microscopy. These aerosol regimes, which can occur in closed, armored military vehicle penetration, are of concern for potential health effects, especially as a consequence of being inhaled. In a controlled volume containing 10 equispaced steel target plates, particulates were systematically collected onto special filters. Filter collections were examined by scanning and transmission electron microscopy (SEM and TEM) which included energy-dispersive (X-ray) spectrometry (EDS). Dark-field TEM identified a significant nanoparticle concentration while EDS in the SEM identified the propensity of mass fraction particulates to consist of Fe and FeO, representing target erosion and formation of an accumulating debris field. Direct exposure of human epithelial cells (A549), a model for lung tissue, to particulates (especially nanoparticulates) collected on individual filters demonstrated induction of rapid and global cell death to the extent that production of inflammatory cytokines was entirely inhibited. These observations along with comparisons of a wide range of other nanoparticulate species exhibiting cell death in A549 culture may suggest severe human toxicity potential for inhaled ballistic aerosol, but the complexity of the aerosol (particulate) mix has not yet allowed any particular chemical composition to be identified.
