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Increased Rho GTPase activity is associated with decreased tumor inhibitory activity of immortalized fibroblasts

ABSTRACT: Clone (Whirly) of human BJhTERT (human foreskin cell line) cells exposed to PC3 mRFP cells Clones (Whirly) of human BJhTERT (human foreskin cell line) cells exposed to PC3 mRFP cells were analyzed with Affymetrix Whole Target (WT) Assay (GeneTitan plate format Gene ST 1.1)

ORGANISM(S): Homo sapiens

SUBMITTER: Hayrettin Guven

PROVIDER: E-GEOD-56832 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Increased Rho GTPase activity is associated with decreased tumor inhibitory activity of immortalized fibroblasts

Project description:Two clones (Whirly and Crossy) of human BJhTERT (human foreskin cell line) cells with different inhibitory capacity were analyzed and compared for their gene expression profile by whole genome microarray. Two clones (Whirly and Crossy) of human BJhTERT (human foreskin cell line) cells were analyzed with Affymetrix Whole Target (WT) Assay (GeneTitan plate format Gene ST 1.1) in order to find out gene expression differences.

2014-03-08 | E-GEOD-55681 | biostudies-arrayexpress

Increased Rho GTPase activity is associated with decreased tumor inhibitory activity of immortalized fibroblasts

2014-03-08 | GSE55681 | GEO

The inhibition of tumor cell proliferation and motility by fibroblasts is both contact and soluble factor dependent

Project description:Clone (Whirly) of human BJhTERT (human foreskin cell line) cells exposed to PC3 mRFP cells

2014-04-17 | GSE56832 | GEO

Expression data from fibroblasts BjhTERT-RhoA-KO and PC3 tumor cells after cocultivation and before

2016-07-01 | GSE83913 | GEO

Expression data from fibroblasts BjhTERT-RhoA-KO and PC3 tumor cells after cocultivation and before

Project description:Gene expression analysis of BJhTERT RhoA-KO and PC3 mRFP was performed before and after six days confrontation in co-cultivation. Analyses of original and control fibroblasts was also performed. To identify molecules that may mediate the tumor promoting effect of RhoA knock out fibroblasts, we performed gene expression analysis of BJhTERT RhoA-KO and PC3 mRFP cells before the confrontation and then after six days after it, using Affymetrix Whole Transcript Assay platform. Four days old fibroblast monolayer was confronted with PC3 mRFP tumor cells, plated in ratio 1:30 to number of plated fibroblasts. After six days of confrontation cells were sorted by FACS (Fluorescence-activated cell sorting). Total RNA was isolated from each confronted and non-confronted cells with kit. Then 150 ng of total RNA were used for transcriptomic analysis.for RNA extraction and hybridization on Affymetrix microarrays

2016-07-01 | E-GEOD-83913 | biostudies-arrayexpress

Rho GTPase activity modulates Wnt3a/beta-catenin signaling.

Project description:Wnt proteins constitute a family of secreted signaling molecules that regulate highly conserved pathways essential for development and, when aberrantly activated, drive oncogenesis in a number of human cancers. A key feature of the most widely studied Wnt signaling cascade is the stabilization of cytosolic beta-catenin, resulting in beta-catenin nuclear translocation and transcriptional activation of multiple target genes. In addition to this canonical, beta-catenin-dependent pathway, Wnt3A has also been shown to stimulate RhoA GTPase. While the importance of activated Rho to non-canonical Wnt signaling is well appreciated, the potential contribution of Wnt3A-stimulated RhoA to canonical beta-catenin-dependent transcription has not been examined and is the focus of this study. We find that activated Rho is required for Wnt3A-stimulated osteoblastic differentiation in C3H10T1/2 mesenchymal stem cells, a biological phenomenon mediated by stabilized beta-catenin. Using expression microarrays and real-time RT-PCR analysis, we show that Wnt3A-stimulated transcription of a subset of target genes is Rho-dependent, indicating that full induction of these Wnt targets requires both beta-catenin and Rho activation. Significantly, neither beta-catenin stabilization nor nuclear translocation stimulated by Wnt3A is affected by inhibition or activation of RhoA. These findings identify Rho activation as a critical element of the canonical Wnt3A-stimulated, beta-catenin-dependent transcriptional program.

| S-EPMC2735600 | biostudies-literature

Regulation of Rho GTPase crosstalk, degradation and activity by RhoGDI1.

Project description:At steady state, most Rho GTPases are bound in the cytosol to Rho guanine nucleotide dissociation inhibitors (RhoGDIs). RhoGDIs have generally been considered to hold Rho proteins passively in an inactive state within the cytoplasm. Here we describe an evolutionarily conserved mechanism by which RhoGDI1 controls the homeostasis of Rho proteins in eukaryotic cells. We found that depletion of RhoGDI1 promotes misfolding and degradation of the cytosolic geranylgeranylated pool of Rho GTPases while activating the remaining membrane-bound fraction. Because RhoGDI1 levels are limiting, and Rho proteins compete for binding to RhoGDI1, overexpression of an exogenous Rho GTPase displaces endogenous Rho proteins bound to RhoGDI1, inducing their degradation and inactivation. These results raise important questions about the conclusions drawn from studies that manipulate Rho protein levels. In many cases the response observed may arise not simply from the overexpression itself but from additional effects on the levels and activity of other Rho GTPases as a result of competition for binding to RhoGDI1; this may require a re-evaluation of previously published studies that rely exclusively on these techniques.

| S-EPMC2866742 | biostudies-literature

Transcription profiling of human multiple myeloma cell lines treated with EZH2 inhibitor E7438 and DMSO controls

Project description:We used a total of 10 human multiple myeloma cell lines for the study (KMS-20, KMS-28BM, MOLP-8, RPMI-8226, U-266, KMS-11, KMS-12-PE, KMS-34, LP-1 and NCI-H929). Cells (2 x 105 per well) were seeded into six-well culture plates in the appropriate culture media 24 h before treatment. Five replicate wells were then exposed to 2 uM of E7438 or control DMSO for 3 days. Cells were collected and RNA was extracted using RNeasy kit (Qiagen) according to the manufacturer’s instructions. For each sample, 250ng of total RNA was amplified using the Affymetrix GeneChip WT PLUS Reagent Kit according to the protocol described in User Manual Target Preparation for GeneChip Whole Transcript (WT) Expression Arrays (P/N 703174 Rev. 2). An Affymetrix Human Gene 2.1 ST 96-array plate was hybridized with 3µg of fragmented and labeled ss cDNA, washed, stained, and scanned according to the protocol described in User Manual GeneTitan®Instrument User Guide for Expression Arrays Plates (P/N 702933 Rev.1) and Affymetrix® GeneChip® Command Console™ User’s Guide (P/N 702569 Rev.9) using the Affymetrix GeneTitan instrument.

2016-01-11 | E-MTAB-3540 | biostudies-arrayexpress

Molecular perturbation strategies to examine spatiotemporal features of Rho GEF and Rho GTPase activity in living cells.

Project description:Much of our current knowledge of Rho GTPase networks and the regulation by Rho guanine exchange factors (Rho GEFs) and Rho GTPase activating proteins (Rho GAPs) is based on population-based techniques. Over the last decades, technologies that enable single cell analysis with high spatial and temporal resolution have revealed that Rho GTPase activity in cells is regulated on second timescales and at submicrometer length scales. Therefore, perturbation methods with matching spatial and temporal resolution are crucial to further our understanding of Rho GTPase signaling. Here, we give a brief overview of the components of Rho GTPase signaling networks and review a range of existing perturbation strategies that target a specific component of the Rho GTPase signaling module. The advantages and limitations of each perturbation method are discussed. Several recommendations are formulated to guide future studies aimed at addressing spatiotemporal aspects of Rho GEF and Rho GTPase signaling.

| S-EPMC6548299 | biostudies-literature

Rho guanine nucleotide exchange factors: regulators of Rho GTPase activity in development and disease.

Project description:The aberrant activity of Ras homologous (Rho) family small GTPases (20 human members) has been implicated in cancer and other human diseases. However, in contrast to the direct mutational activation of Ras found in cancer and developmental disorders, Rho GTPases are activated most commonly in disease by indirect mechanisms. One prevalent mechanism involves aberrant Rho activation via the deregulated expression and/or activity of Rho family guanine nucleotide exchange factors (RhoGEFs). RhoGEFs promote formation of the active GTP-bound state of Rho GTPases. The largest family of RhoGEFs is comprised of the Dbl family RhoGEFs with 70 human members. The multitude of RhoGEFs that activate a single Rho GTPase reflects the very specific role of each RhoGEF in controlling distinct signaling mechanisms involved in Rho activation. In this review, we summarize the role of Dbl RhoGEFs in development and disease, with a focus on Ect2 (epithelial cell transforming squence 2), Tiam1 (T-cell lymphoma invasion and metastasis 1), Vav and P-Rex1/2 (PtdIns(3,4,5)P3 (phosphatidylinositol (3,4,5)-triphosphate)-dependent Rac exchanger).

| S-EPMC4875565 | biostudies-other

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