Gene regulation in human epidermal keratinocytes by a Rho kinase inhibitor (Y-27632)
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE52515 | GEO | 2014/03/31
SECONDARY ACCESSION(S): PRJNA229149
REPOSITORIES: GEO
Similar Datasets
Project description:Keratinocytes are a targeted cells for infection of human papillomaviruses (HPV). But, there are very limited information for gene regulation of human karatinocytes by HPV infection. Y-27632, a rho-kinase inhibitor, stimulates proparation of keratinocytes. We identified genes, which are regulated by a drug for karatinocyte proparation (Y-27632). mRNA profiles of primary HEKs regulated by a rho kinase inhibitor (Y-27632, Y-drug) were generated by microarray analysis using Illumina BeadChip.
2014-03-31 | E-GEOD-52515 | biostudies-arrayexpress
Project description:Keratinocytes are the most abundant cell type in the epidermis. They prevent desiccation and provide immunological and barrier defense against potential pathogens such as Staphylococcus aureus and Candida albicans. The study of this first line of immune defense may be hindered by invasive isolation methods and/or improper culture conditions to support stem cell maintenance and other potential mechanisms contributing to long-term subcultivation in vitro. Primary keratinocytes have been successfully isolated from blister roofs induced by negative pressure, which separates the epidermis from the dermis in vivo in human subjects. This method allows collection of pure epidermal cells without dermal contamination in a minimally invasive manner. However, the isolated keratinocytes differentiate and senesce when cultured in vitro beyond five passages. Here, we present evidence that the Rho kinase (ROCK) inhibitor Y-27632 can be used to effectively increase the proliferative capabilities of keratinocytes isolated using the suction blister method, similar to what has been previously reported for primary keratinocytes isolated using alternative methods. We show that the increase in passage number is directly correlated to delayed differentiation, and that cells passaged long term with the inhibitor retain their ability to stratify in organotypic raft cultures and respond to cytokine treatment; additionally, the late passage cells have a heterogeneous mix of differentiated and non-differentiated cells which may be predicted by a ratio of select differentiation markers. The described method presents a minimally invasive procedure for keratinocyte isolation and prolonged culture that allows analysis of keratinocyte function in both healthy volunteers and patients with dermatologic diseases.
2018-09-01 | GSE119317 | GEO
Project description:The study tried the identify some differentially expressed proteins in rat lingual keratinocytes treated with or without the ROCK inhibitor Y-27632. In brief, rat lingual keratinocytes were treated with or without Y-27632 for 36 h and were subjected to proteomic analysis.
2022-06-23 | PXD025812 | JPOST Repository
Project description:Prolonging culture of primary human keratinocytes isolated from suction blisters with the Rho kinase inhibitor Y-27632
| PRJNA488732 | ENA
Project description:Expression data from this experiment is part of a larger project aimed at defining the individual effects and synergistic effects of ROCK inhibitor Y-27632 and conditioned media from irradiated J2 cells when applied to epithelial cells. This data set consists of four individual samples, each of which are total RNA collected from human foreskin keratinocyte cells, either grown in F medium (control), treated with Y-27632, grown in conditioned medium (as described in associated publication), or both treatments.
2014-09-15 | GSE61226 | GEO
Project description:Microarray expression data from three different cell lines of human embrionic stem cells (Hs181, Hs293, Hs420), grown under feeder and feeder-free conditions (matrigel) with the addition of Y-27632 (ROCK Inhibitor) We assessed the difference between cell lines and the impact of feeder-free vs matrigel conditions upon the addition of the ROCK Inhibitor Y-27632
2018-01-01 | GSE102108 | GEO
Project description:Purpose: Y-27632, a well-characterized Rho-associated kinase (ROCK) inhibitor, has been shown to play an important role in regulation of cell growth in multiple types of cells, the regulation appeared to be cell specific. To further understand its molecular mechanisms, we performed RNA-seq analysis of HDF at 12 hours and 48 hours after Y-27632 treatment. Methods: Total RNAs were isolated from dermal cells with/without Y-27632 treatment for 12 hours or 48 hours respectively. Then mRNAs were purified from above total RNA using poly-T oligo-attached magnetic beads. After sequencing libraries were prepared using NEBNext® UltraTM RNA Library Prep Kit for Illumina® (NEB, USA), the library preparations were sequenced on an Illumina Hiseq platform and 125 bp/150 bp paired-end reads were generated. Results: We sequenced 12 of 229 differential expressed genes 48 hours after Y-27632 treatment and found genes which by GO analysis are negative regulators of cell growth ). Of these 12 genes, the expression level of 4 genes, IGFBP1, IGFBP-5, HNF4A and NKD1, changed significantly at both 12 hours and 48 hours of Y-27632 treatment. This results, validated by RT-qPCR analysis showed that the expression of IGFBP-5 (Insulin Growth Factor Binding Protein 5) increased by 1.5 fold after 12 hours of treatment, and increased more than 10-fold after 48 hours. Expression of the other 3 genes didn’t change significantly after Y-27632 treatment Conclusions:RNA-seq analysis of gene expression profile in HDFs collected from 12 hours and 48 hours treatment of Y-27632. From these analyses we found the IGFBP-5, a negative regulation of cell proliferation, whose expression increased with the duration of Y-27632 treatment (7-fold increase after the first 24 hours
2019-08-19 | GSE135934 | GEO
Project description:We have used microarray technology to identify the transcriptional targets of Rho subfamily GTPases. This analysis indicated that murine fibroblasts transformed by these proteins show similar transcriptomal profiles. Functional annotation of the regulated genes indicate that Rho subfamily GTPases target a wide spectrum of biological functions, although loci encoding proteins linked to proliferation and DNA synthesis/transcription are up-regulated preferentially. Rho proteins promote four main networks of interacting proteins nucleated around E2F, c-Jun, c-Myc, and p53. Of those, E2F, c-Jun and c-Myc are essential for the maintenance of cell transformation. Inhibition of Rock, one of the main Rho GTPase targets, leads to small changes in the transcriptome of Rho-transformed cells. Rock inhibition decreases c-myc gene expression without affecting the E2F and c-Jun pathways. Loss-of-function studies demonstrate that c-Myc is important for the blockage of cell-contact inhibition rather than for promoting the proliferation of Rho-transformed cells. However, c-Myc overexpression does not bypass the inhibition of cell transformation induced by Rock blockage, indicating that c-Myc is essential, but not sufficient, for Rock-dependent transformation. These results reveal the complexity of the genetic program orchestrated by the Rho subfamily and pinpoint protein networks that mediate different aspects of the malignant phenotype of Rho-transformed cells Keywords: Rho/Rac GTPases, microarray, oncogenesis, proliferation, gene expression, transcription, Rock, c-Myc
2007-01-03 | GSE5913 | GEO
Project description:Cellular differentiation is regulated through activation and repression of defined transcription factors. A hallmark of differentiation is a pronounced change in cell shape, which is determined by dynamics of the actin cytoskeleton. In de-differentiated fat (DFAT) cells and 3T3-L1 cells, we showed that treatment with the ROCK inhibitor Y-27632, by inducing remodeling of the actin cytoskelton, causes adipocyte differentiation. In addition, we found that depletion of MKL1, an actin binding transcriptional coactivator, elicits adipogenesis. To investigate whether regulation of MKL1 by actin cytoskeleton dynamics drives adipocyte differentiation, we compared the gene expression changes resulting from DFAT cells after treatment with Y-27632 and transfection of Mkl1 siRNA. Using Affymetrix mouse genome array, we compared global gene expression profiles in DFAT cells after treatment with Y-27632 and transfection with Mkl1 siRNA to search for particular biological functions in genes of which expression intensities were increased and decreased.
2014-03-28 | E-GEOD-52334 | biostudies-arrayexpress
Project description:After thawing, the mechanism of apoptosis advancing in human ES cells was invesitigated mainly by Gene conpath analysis. As time passed after thawing, nuclear fragmented cells increased more and more in cR-/mR- and cR+/mR-, but in cR+/mR+ and cR-/mR+, thawed cells formed cluster markedly, which was seen among three cell lines. Flow cytometry using TUNEL assay supported final colony formation ratio, which defined that such cell death was due to apoptosis. Gene map analysis 12 hours later after thawing of cR-/mR- showed activation of IL-1? & its receptor IL-1R and TGF-? & its receptor ACVR1C required for activation of caspase-8, initiation of caspase-8 and 10 and marked activation of ARHGDIB promoting actin reorganization comparing to cR+/mR+. Recovered cell lines in any condition did not lose proliferation and pluripotency regardless of ROCK inhibitor treatment.?These results showed apoptotic events after thawing were more marked in cR- and advanced through autoenhanced cascaded mechanism followed by initiation of self-cytokain activity, and were suppressed by ROCK inhibitor Y-27632. To investigate the protective mechanism caused by a novel cryo-technique of human embryonic stem (hES) cells using ROCK inhibitor Y-27632 against the cell death due to cryopreservation and thawing with Gene map analysis in addition to estimations of survival rate, apoptosis, undifferentiated states and pluripotency, the dissociated hES cells were cryopreserved in a freezing container in the following four conditions: addition of 10µM of Y-27632 to 1: cryoprotection solution and post-thawing medium (cR+/mR+), 2: only addition to cryoprotection (cR+/mR-), 3: only addition to post-thawing medium (cR-/mR+), 4: no addition to any medium (cR-/mR-).
2011-11-15 | E-GEOD-21796 | biostudies-arrayexpress