Project description:REDD1 KO affects basal global gene expression profile and transcriptomic response to wounding and glucocorticoid treatment in HaCaT human keratinocytes
Project description:New alkyl-phospholipids (APLs) that are structurally derived from the platelet-activating factor are promising candidates for anticancer treatment. After the incorporation into cell membranes, APLs are able to interfere with a wide variety of key enzymes implicated in cell growth, motility, invasion and apoptosis. Besides the prototype edelfosine, we presented a novel group of APLs, glycosidated phospholipids that efficiently inhibit cell proliferation. Two members of this group, Ino-C2-PAF and Glc-PAF, display high efficacy and low cytotoxicity in immortalized non-tumorigenic skin keratinocyte cell line HaCaT. However, the influence of APLs on the transcription of the whole genome is still unknown. Here, using Agilent cDNA microarray technology, we compared global gene expression profiles of HaCaT cells treated with edelfosine, Ino-C2-PAF or Glc-PAF with the profile of control cells. The influence of APLs on the transcriptional profile of immortalized keratinocytes (HaCaT) was analyzed treating HaCaT cells with respectively 5 M-BM-5M Ino-C2-PAF, Glc-PAF and edelfosine for 24 h. Control cells were left untreated. Three independent experiments were performed for each condition.
Project description:Immortalized keratinocytes HaCaT is popular model for skin research (toxicity, irritation, allergic reactions or interaction of cells). They maintain stable keratinocyte phenotype and respond to keratinocyte differentiation stimuli. However, the programs of stratification and expression of differentiation markers in HaCaT keratinocytes are aberrant. HaCaT cells bear two mutant p53 alleles (R282Q and H179Y) which contain Gain-of-Function (GOV) mutations acquired as a result of spontaneous immortalization (mutp53). At the same time, mutp53 acts as a transcription factor, and also affects interaction of p63 protein with its transcription targets. It is known that proteins of the P53 family play an important role in regulating processes of proliferation and differentiation of human keratinocytes.At the same time, the role of mutp53 in these processes is not fully clear. We present data sets obtained as a result of high-performance proteomic analysis of immortalized HaCaT keratinocytes with p53 knockout in two different states: sub-confluent and confluent, which are characterized by different intensity of cell differentiation processes. 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.)
Project description:SWATH-MS data for human keratinocytes (HaCaT cell line), sample 010-012 indicates HaCaT cells with no treatment, sample 013-015 indicates HaCaT cells induced by erastin, sample 0016-018 indicates HaCaT cells treated by erastin and CBD. For more detailed information, please contact Dr.Chang Liu (hichang813@uri.edu)
Project description:To figure out the expression level of genes in normal human keratinocytes, we performed RNA-seq analysis in HaCaT cells. Here, we used 4 HaCaT cells as parallel samples.
Project description:SWATH-MS data for human keratinocytes (HaCaT cell line), sample 001-004 indicates HaCaT cells with no treatment, sample 005-008 indicates HaCaT cells induced by erastin, sample 009-012 indicates HaCaT cells induced by erastin plus MGO (methyglyoxal), and sample 013-016 indicates HaCaT cells induced by erastin plus GO (glyoxal). For more detailed information, please contact Dr.Chang Liu (hichang813@uri.edu)
Project description:In an RNAseq analysis, we have identified the microRNA hsa-miR-129-5p with high levels in acute wounds (day1 to day7) compared to normal skin. The biological function of this miRNA in human epidermal keratinocytes during wound repair has not been studied. To study the genes regulated by miR-129-5p , we transfected miR-mimics targeting miR-129-5p into human primary epidermal keratinocytes to over-express miR-129-5p expression. We performed a global transcriptome analysis of keratinocytes upon miRNA overexpression using Affymetrix arrays.
