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Human keratinocytes HaCaT LC-MSMS_SDS exposure

ABSTRACT: We employed human HaCaT cells as a model system to identify cellular proteins that accompany SDS-induced toxicity based on a proteomic approach. HaCaT human keratinocyte cell line were treated with a non-cytotoxic dose of SDS (25 µg/ml, as determined by the MTT assay and microscopically examination) for 48 h. The altered abundance of proteins from HaCaT keratinocytes exposed to SDS was analyzed by LC-MS/MS approach and quantified using Progenesis LC software. The abundance of 217 proteins (which were identified by multiple peptides, ≥ 2) was altered in keratinocytes exposed to SDS; in which 131 proteins had increased abundance while 86 proteins was down regulated. The Pathview map of 131 up-regulated proteins was built and enhancement of glycolysis/gluconeogenesis was found.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Keratinocyte

SUBMITTER: Natalia Petushkova

LAB HEAD: Andrey V. Lisitsa

PROVIDER: PXD007789 | Pride | 2017-11-03

REPOSITORIES: Pride

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			Action	DRS
	170616ONA_07.mgf	Mgf
	170616ONA_07.raw	Raw
	170616ONA_07_160622035546.mgf	Mgf
	170616ONA_07_160622035546.raw	Raw
	170616ONA_07_160622052741.mgf	Mgf

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Publications

[Proteomic profiling of HaCaT keratinocytes exposed to skin damaging detergents].

Rusanov A L AL Petushkova N A NA Poverennaya E V EV Nakhod K V KV Larina O V OV Lisitsa A V AV Luzgina N G NG

Biomeditsinskaia khimiia 20171001 5

The effects of sodium dodecyl sulfate (25 mg/ml) and Triton X-100 (12.5 mg/ml and 25 mg/ml) on the HaCaT immortalized keratinocytes exposed to these surfactants for 48 h were studied. Using shotgun proteomics, a comparative analysis of the proteomic profiles of control and experimental cells after surfactants exposure was carried out. 260 common proteins were identified in control and experimental cells; 33 proteins were found in cells exposed to all three treatments, but not in control cells. T ...[more]

PMID: 29080872

Similar Datasets

Human keratinocytes HaCaT LC-MS/MS

Project description:Triton X-100 has been used as a model substance to study the effects of irritants on the skin. HaCaT human keratinocyte cell line were treated with a non-cytotoxic dose of Triton X-100, as determined by the MTT assay and microscopically examination) for 48 h. The altered abundance of proteins from HaCaT keratinocytes exposed to Triton X-100 was analyzed by LC-MS/MS approach and quantified using Progenesis LC software.

2017-11-03 | PXD007776 | Pride

Proteome profiling of HaCaT keratinocytes

Project description:An important step in the proteomic analysis is the optimal extraction and the processing of protein material in order to ensure the highest sensitivity in downstream protein detection. Cell lysis is important for proteomic profiling with an eventual aim to discover differently expressed proteins (DEPs). Herein for cell lysis, two sample preparation protocols were compared: 0.2% SDS-based solubilization combined with the 1DE-gel concentration (Protocol 1) and osmotic shock (Protocol 2). HaCaT keratinocytes were cultured with/without sodium dodecyl sulfate (SDS) and used to test the hypothesis that SDS might be associated with the initiation and development of malignancies. Protocol 1 increased sensitivity and coverage of HaCaT proteome, it permitted us to reveal the provided new insights into the skin impact of SDS (widely used and rather non-toxic component of household chemicals, cosmetic products, shampoos and so on). Our results suggest the possible role of SDS in skin cancerogenesis.

2022-07-20 | PXD035202 | Pride

Aberrantly expressed genes in HaCaT keratinocytes chronically exposed to arsenic trioxide

Project description:The purpose of this study is to search for aberrant genes in HaCaT keratinocytes after chronic exposure to arsenic trioxide. The objective of the investigation was to discover the mechanism of arsenic carcinogenicity in human epidermal keratinocytes. We hypothesize that a combined strategy of DNA microarray, qRT-PCR and gene function annotation will identify aberrantly expressed genes in HaCaT keratinocyte cell line after chronic treatment with arsenic trioxide. HaCaT cells were chronically exposed to 0.5M-BM-5g/mL arsenic trioxide (As2O3) up to 22 passages and RNA was extracted. Microarray data analysis identified 14 up-regulated genes and 21 down-regulated genes in response to arsenic trioxide Two experimental groups: 1. The treatment group was sub-cultured up to passage 22 to establish a chronic exposure state. 2. The passage control group was also sub-cultured up to 22 passages but with no exposure to arsenic trioxide. 4 technical replicates with 3 replicates making a total of 8X3 =24 samples HaCat Cell untreated (passage control): 1. H1_H001, H1_H002, H1_H003 2. H2_ H004, H2_H005, H2_H006 3. H3_ H007, H3_H008, H3_H009 4. H4_ H010, H4_H011, H4_H012 HaCat Cell treated with 0.5M-BM-5g/ml of arsenic trioxide: 5. A1_H013, A1_H014, A1_H015 6. A2_H016, A2_H017, A2_H018 7. A3_H019, A3_H020, A3_H021 8. A4_H022, A4_H023, A4_H024 Cell Type: Human Skin Keratinocyte: 1.5 M-CM-^W105 HaCaT cells were cultured in 7.5 ml of complete DMEM containing 10% Fetal Bovine Serum (FBS) and 1% penicillin, streptomycin in T-25 culture plate. Cells were incubated in a humidified atmosphere with 5% CO2 at 37 M-BM-:C. The treatment groups were exposed to 0.5M-BM-5g/mL As2O3 (equivalent to LC 0.5), and passaged at 90% confluent. Total RNA was extracted from 4 technical replicates of unexposed HaCaT cells and HaCaT cells chronically exposed to arsenic trioxide up to passage 22 using RNA STAT-60 (TEL-TEST, INC, Friendswood, TX, USA).

2012-04-11 | E-GEOD-37192 | biostudies-arrayexpress

Transcription profiling of human ES and keratinocyte cells in response to ionising radiation

Project description:I-131, a source of beta- and gamme radiation and the Auger electron emitter I-125 dU incorporated into nuclear DNA were compared in two cell lines , human ES cells and the keratinocyte cell line HaCaT.

2010-10-07 | E-MEXP-1843 | biostudies-arrayexpress

The impact of the HSPA2 gene knockout on transcriptome profile in immortalized epidermal keratinocytes (HaCaT line)

Project description:The specific functional features of the epidermal keratinocytes are determined by the activity of many genes. The aim of the project was to characterize the role of HSPA2, a member of the HSPA chaperone family (HSP70), in human epidermal keratinocytes. The inactivation of the HSPA2 gene in the HaCaT line of spontaneously immortalized epidermal keratinocytes was performed by CRISPR/Cas9 gene editing system. Next, the effect of modifications on the transcriptomic profile of cells growing in a three-dimensional model of reconstructed human epidermis in vitro was investigated. The cells were grown at air-liquid interface culture on collagen-fibroblast matrix to achieve maximal level of HaCaT differentiation in RHE system.

2023-09-25 | E-MTAB-13166 | biostudies-arrayexpress

The impact of the HSPA2 gene downregulation on transcriptome profile in immortalized epidermal keratinocytes (HaCaT line) grown in 3D cell culture

Project description:The specific functional features of the epidermal keratinocytes are determined by the activity of many genes. The aim of the project was to characterize the role of HSPA2, a member of the HSPA chaperone family (HSP70), in human epidermal keratinocytes. The downregulation of HSPA2 gene in HaCaT cell line was performed by lentivirus-mediated shRNA method. Next, the effect of modifications on the transcriptomic profile of cells growing in a three-dimensional model of reconstructed human epidermis in vitro was investigated. The cells were grown at air-liquid interface culture on extracellular matrix to achieve maximal level of HaCaT differentiation in RHE system. This study was supported by a Polish National Science Center grant number NCN 2017/25/B/NZ4/01550.

2023-11-29 | E-MTAB-13266 | biostudies-arrayexpress

Study of transcriptional effects of monosodium urate crystal (MSU) on HaCaT keratinocyte cell line.

Project description:Aim of the study was to characterize at a molecular level (changes in transcriptomes) the effect of monosodium urate crystal (MSU) on HaCaT keratinocyte cell line. This was adressed by using a culture model. The HaCaT cell line (human keratinocytes) was stimulated by MSU (1mg/mL) vs control for 12 hrs. By using genome-wide expression profiling, we identified deregulation of functionally relevant gene networks. HaCaT were obtained from Cell Lines Service (Eppelheim, Germany) and grown in DMEM medium (PAN biotech, Aidenbach, Germany) supplemented with 10% FBS (Life Technology, Grand Island, NY, USA), L-glutamine and non-essential amino acid. Before the treatment HaCaT cells were cultured in serum-free medium for 12hrs. HaCaT were treated with MSU (1mg/ml) vs DMEM control for 12hrs then submitted to RNA extration and gene expression profiling. Triplicate experiments were performed: HaCaT control (n=3), MSU-treated (n=3).

2015-10-29 | E-GEOD-65931 | biostudies-arrayexpress

Human keratinocyte sensitizing potential

Project description:HaCaT human keratinocytes were exposed for 4 hours to 26 compounds at a concentration that resulted in 20% cell death after 24 hours of exposure.

2012-12-04 | E-MTAB-943 | biostudies-arrayexpress

Deciphering Staphylococcus aureus-host dynamics using dual Activity-based protein profiling of ATP-interacting proteins

Project description:This study focuses on ATP-binding proteins to dissect the complex interplay between Staphylococcus aureus and human cells, particularly macrophages (THP-1) and keratinocytes (HaCaT), during the course of an intracellular infection. A snapshot of the various protein activity and function is provided using a desthiobiotin-ATP probe, which targets ATP-interacting proteins. In S. aureus, we observe enrichment in pathways required for nutrient acquisition, biosynthesis and metabolism of amino acids and energy metabolism when located inside human cells. Additionally, the direct profiling of the protein activity revealed specific adaptations of S. aureus to the keratinocytes and macrophages. Mapping the differentially activated proteins to biochemical pathways in the human cells with intracellular bacteria revealed cell-type specific adaptations to bacterial challenges where THP-1 cells prioritized immune defenses, autophagic cell death, and inflammation. In contrast, HaCaT cells emphasized barrier integrity and immune activation. We also observe bacterial modulation of host processes and metabolic shifts.

2024-04-22 | PXD049003 | Pride

Expression data from BCL-3 overexpression HaCat cells

Project description:Constitutive activation of EGFR- and NF-kB-dependent pathways is a hallmark of cancer, yet signaling proteins that connect both oncogenic cascades are poorly characterized. Here we define KIAA1199 as a BCL-3- and p65-dependent gene in transformed keratinocytes. KIAA1199 expression is enhanced upon human papillomavirus (HPV) infection and is aberrantly expressed in clinical cases of cervical (pre)neoplastic lesions. Mechanistically, KIAA1199 binds Plexin A2 and protects from Semaphorin 3A-mediated cell death by promoting EGFR stability and signaling. Moreover, KIAA1199 is an EGFR-binding protein and KIAA1199 deficiency impairs EGF-dependent Src, MEK1 and ERK1/2 phosphorylations. Therefore, EGFR stability and signaling to downstream kinases requires KIAA1199. As such, KIAA1199 promotes EGF-mediated epithelial-mesenchymal transition (EMT). Taken together, our data define KIAA1199 as an oncogenic protein induced by HPV infection and constitutive NF-kB activity that transmits pro-survival and invasive signals through EGFR signaling. We used microarrays to detail the global programme of gene expression upon BCL-3 overexpression We used two experimental conditions, namely HaCat cells infected with a control lentivirus as well as HaCat cells infected with a BCL-3 expressing construct. Both experimental conditions were in triplicates.

2014-11-04 | E-GEOD-60551 | biostudies-arrayexpress

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