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Effect of cell density on HaCaT keratinocyte gene expression

ABSTRACT: Effect of cell density on HaCaT keratinocyte gene expression

PROVIDER: PRJNA1040225 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR26826567_1.fastq.gz	Fastqsanger.gz
	SRR26826567_2.fastq.gz	Fastqsanger.gz
	SRR26826568_1.fastq.gz	Fastqsanger.gz
	SRR26826568_2.fastq.gz	Fastqsanger.gz
	SRR26826569_1.fastq.gz	Fastqsanger.gz

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Effect of cell density on HaCaT keratinocyte gene expression

Project description:Cell density affects keratinocyte behaviors and gene expression. Here, We examines the differential expressed genes regulated by cell density. We performed RNA-seq analysis to compare HaCaT cells cultured under high- or low0density conditions. The Gene Ontology Biological Process terms enriched among differential expressed genes through DESeq2 included cell adhesion and keratinocyte differentiation. Our data provides the understanding of keratinocyte behavior regulated by cell density.

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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

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

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The miRNA microarray data after UVB exposure to human keratinocyte HaCaT cells

Project description:UVB plays a key role in inflammation and DNA damage in human skin. Human keratinocyte HaCaT cells were utilized to determine the up- and down-regulated miRNA after UVB exposure. We used microarrays to identify the miRNA affected after UVB (15 mJ/cm^2) exposure to HaCaT cells

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HaCaT keratinocytes day3 vs HaCaT keratinocytes day10

Project description:The keratinocyte cell line HaCaT was cultured for three days (proliferation) or for ten days (differentiation). RNA from cells at day3 was compared to RNA from cells at day10. The microarray hybridizations were performed in dye-swap procedure : RNA from cells at day3 was labeled with Cy3 (GSM4674, GSM4675, GSM4682, GSM4683) and then with cy5 (GSM4680, GSM4681). Keywords = cell density differentiation program in human keratinocytes

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Extremely low frequency electromagnetic fields (ELF-EMFs) effect on the genetic expression profile of a human keratinocyte line (HaCaT)

Project description:Clinical evidence has shown that electromagnetic fields produce a benefical therapeutic result for wounds, however little is still known about their exact mechanism of action. Moreover, clinical results concerning skin tissue restoration are still debated. In the present study, we carried out gene expression profiling of a human keratinocyte line (HaCaT) submitted to 1 hour of ELF-EMF (frequency 50Hz, intensity 1 mT) in order to identify up- and downregulated genes by this stimulus, and to verify the presence of specific molecular pathways activation. Most of the genes modulated were involved in mechanisms such as protein synthesis. In particular, these genes are related to Mammalian target of Rapamycin (mTOR), which has been identified as a kinase with a pivotal role in cellular proliferation and survival in mammals. In this study, we analyzed the expression profiles of 3 biological replicates of HaCaT cells exposed to ELF-EMF (frequency 50Hz, intensity 1 mT) for 1 hour. All HaCaT cells exposed to ELF-EMF RNAs were hybridized against control sham-exposed HaCaT cells RNAs. Each biological sample was repeated with a technical replicate (extraction-labeling) and a dye-swap experiment.

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Project description:The p53 protein is encoded by TP53 gene and plays the key role in significant number of cellular processes including proliferation, apoptosis and regulation of many stress response pathways. P53 acts like a direct transcription activator of numerous genes regulating cell cycle arrest, DNA repair, growth inhibition and many others (Mollereau and Ma, 2014). The canonical biological function of p53 is maintaining genome integrity via elimination of damaged or exposed to genotoxic stress cells. Immortalized HaCaT cells are widely used for keratinocyte research, since they maintain stable keratinocyte phenotype, have nearly unlimited proliferative potential, do not require specific growth and differentiation factors (Colombo et al., 2017). Also, HaCaT cells produce typical differentiation markers such as cytokeratins K14 and K10, involucrin (Colombo et al., 2017) and respond to keratinocyte differentiation stimuli. Taking together, HaCaT cells have similar to normal human keratinocytes (NHK) properties, however, as many of spontaneously immortalized cell lines HaCaT cells bear two mutant p53 alleles - R282Q and H179Y (Lehman et al., 1993). Mutp53 in HaCaT has an increased affinity to other p53 family members (p63, p73), which significantly expands p53 properties. Moreover, mutp53 indirectly affects specific target genes via protein-protein interactions with other transcription factors (NF-Y, E2F1, NF-KB) or by tethering p63 to new promotor locations. For more detailed investigation of mutp53 impact on various processes in HaCaT cells we performed a shRNA mediated knockdown of mutp53. For generation of stable TP53 knockdown we employed plasmid vector pLKO-p53-shRNA-941 (Addgene # #25637) followed by puromycin selection of transduced cells. Here we present proteomic dataset obtained from wild type HaCaT cells and p53 knock down HaCaT keratinocytes.

2022-04-28 | PXD033538 | Pride

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