Comparative proteoinformatics revealed the essentials of SDS impact on HaCaT keratinocytes

Abstract

There is no direct evidence supporting that SDS is a carcinogen, so to investigate this fact, we used HaCaT keratinocytes as a model of human epidermal cells. To reveal the candidate proteins and/or pathways characterizing the SDS impact on HaCaT, we proposed comparative proteoinformatics pipeline. For protein extraction, the performance of two sample preparation protocols was assessed: 0.2% SDS-based solubilization combined with the 1DE-gel concentration (Protocol 1) and osmotic shock (Protocol 2). As a result, in SDS-exposed HaCaT cells, Protocol 1 revealed 54 differentially expressed proteins (DEPs) involved in the disease of cellular proliferation (DOID:14566), whereas Protocol 2 found 45 DEPs of the same disease ID. The ‘skin cancer’ term was a single significant COSMIC term for Protocol 1 DEPs, including those involved in double-strand break repair pathway (BIR, GO:0000727). Considerable upregulation of BIR-associated proteins MCM3, MCM6, and MCM7 was detected. The eightfold increase in MCM6 level was verified by reverse transcription qPCR. Thus, Protocol 1 demonstrated high effectiveness in terms of the total number and sensitivity of MS identifications in HaCaT cell line proteomic analysis. The utility of Protocol 1 was confirmed by the revealed upregulation of cancer-associated MCM6 in HaCaT keratinocytes induced by non-toxic concentration of SDS. Data are available via ProteomeXchange with identifier PXD035202.

Silver-nanoparticle-modified nanocellulose synthesized by pyroligneous acid: cytotoxicity towards HaCat cells

“…Herein, we set out to discovering the conditions under which the NC-AgNPs toxicity emerges. Due to their high capacity to differentiate and proliferate in vitro, human skin keratinocyte cells (HaCaT) have been extensively utilized in biology and differentiation research [35]. To check the toxicity of NC-AgNPs, HaCaT cells were exposed to NC-AgNPs at different concentrations and intervals, with results presented in figures 6 and 7.…”

Exploring the Functions of Mutant p53 through TP53 Knockout in HaCaT Keratinocytes

“…HaCaT keratinocytes, a line of spontaneously immortalized human keratinocytes, represent a convenient model for research in this field [5]. Although HaCaT cells are traditionally defined as normal non-tumoral keratinocytes and are commonly used as an alternative to primary keratinocytes [6][7][8], they also bear two UV-signature gain-of-function (GOF) mutations in both alleles of TP53 (R282Q and H179Y) [9]. The novel properties of p53 obtained through mutations are manifested in its capacity to bind atypical response elements and its altered affinity to other transcription factors, which in turn results in the suppression of their biological functions [10].…”