RESUMO
Using the method of shotgun mass spectrometry, we have evaluated changes in the proteomic profile of HaCat cells in response to the treatment with sodium dodecyl sulfate (anionic surfactant) and Triton-X100 (non-ionic surfactant) in two concentrations (12.5 µg/ml and 25.0 µg/ml). The study revealed induction of orphan CYP2S1 (biotransformation phase I) in response to Triton-X100. We have identified proteins of II (glutathione-S-transferases, GSTs) and III (solute carrier proteins, SLCs) biotransformation phases, as well as antioxidant proteins (peroxiredoxins, PRDXs; catalase, CAT; thioredoxin, TXN). Thus, proteins of all three xenobiotic detoxification phases were detected. The presented results suggest a new prospect of using HaCaT keratinocytes as a model of human epidermis for studying the metabolism of drugs/toxicants in human skin in vitro.
Assuntos
Proteômica , Tensoativos , Humanos , Tensoativos/farmacologia , Queratinócitos , Linhagem Celular , Pele , Octoxinol , Sistema Enzimático do Citocromo P-450RESUMO
Learning of the molecular mechanisms of the pathological processes development in the normal human keratinocytes (NHK) are difficult. Immortalized keratinocytes HaCaT are often used as an analogue of NHK since they have a number of advantages over the latter - they do not require the presence of growth and differentiation factors in the medium, have unlimited potential for proliferation, demonstrate stable phenotype regardless of the number of passages [1]. Taking into account the properties and characteristics of the HaCaT line, these cells can be considered as a promising experimental model for research of various physiological processes occurring in human keratinocytes. However, to understand the limitations of such an experimental model, a detailed comparative characterization of HaCaT and NHK is required, which can be obtained by carrying out its proteomic analysis. In this article we present datasets obtained through the high-throughput shotgun proteomics analysis of normal human keratinocytes and immortalized HaCaT keratinocytes. As a protocol for proteomic profiling of cells, we used the approach of obtaining LC-MS / MS measurements followed by their processing with Progenesis LC-MS software (Nonlinear Dynamics Ltd.). The mzML files were deposited to the Mendeley Data.
RESUMO
The paper characterizes phenotypic features of subpopulations of human adipose tissue mesenchymal stem cells (MSC) resulting from exposure to Toll-like receptors ligands. MSC1 and MSC2 phenotypes were induced by exposure to LPS and polyinosinic-polycytidylic acid, respectively. Different exposures to Toll-like receptors ligands, short-term (3 h) and long-term (24 h), were studied. The cytokine profile of cells with the MSC2 phenotype differed depending on the duration of exposure to the inductor, while the cytokine profile of cells with the MSC1 phenotype remained stable. Morphometric features of mesenchymal stem cells with the MSC1 and MSC2 phenotypes, as well as differences in the IDO gene expression were identified. These differences can be used to distinguish between these cell types.