Effect of CRISPR-Cas9 mediated knock-out of integrin alpha2 on the transcriptome of DU145 prostate cancer cell grown as a spheroid culture
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE111507 | GEO | 2018/08/02
REPOSITORIES: GEO
Similar Datasets
Project description:Effect of CRISPR-Cas9 mediated knock-out of integrin alpha2 on the transcriptome of DU145 prostate cancer cell grown as a spheroid culture
| PRJNA437244 | ENA
Project description:Previous reports demonstrate that the α2-integrin (α2) mediates pancreatic ductal adenocarcinoma (PDAC) cell interaction with collagens. We found that untransformed pancreatic ductal epithelial cells and well-differentiated PDAC cells use α2 exclusively to adhere and migrate on collagenI. In contrast, poorly-differentiated PT45P1 and MIAPaCa2 cells demonstrate reduced reliance on, or complete loss of α2, respectively. Further, well-differentiated PDAC lines exhibit reduced in vitro invasion compared to poorly-differentiated lines, and α2-blockade suppressed invasion of well-differentiated lines exclusively. Based on these data and the demonstrated role of α2 in maintaining tissue architecture in other organs, we hypothesized that α2 may actually suppress the malignant phenotype in PDAC. Accordingly, stable ectopic expression of α2 in MIAPaCa2 cells (MP2-α2) retarded in vitro invasion. MP2-α2 cells maintained on collagenI were more invasion-retarded than MP2-α2 cells maintained in standard tissue culture, and demonstrated higher α2β1 expression that was reflected in faster and more complete adhesion/migration on collagenI. Affymetrix gene expression profiling of α2-expressing and mock-transfected cells revealed that kallikrein-related peptidases (KLK)-5, 6 and 7 were specifically upregulated by α2. Accordingly, well-differentiated PDAC lines express KLK-5, 6 and 7 and KLK blockade increased invasion as well as collagenI-migration in KLK-positive lines. Importantly, an α2 cytoplasmic deletion mutant promoted KLK expression and retarded invasion, while an α9α2 chimera retarded invasion less efficiently, and did not impact KLK expression. These data demonstrate for the first time that the α2-ectodomain and KLKâ??s coordinately regulate a less invasive phenotype in PDAC cells. Experiment Overall Design: Affymetrix global gene arrays were used to analyse differences in gene expression patterns in MIAPaCa2 cells stably reexpressing the alpha2 integrin, a cytoplasmic deletion of the alpha2 integrin (dCYTO) or an alpha9 ectodomain and transmembrane domain/alpha2 cytoplasmic domain chimera, versus vector-only mock controls. RNA was harvested from cells passaged identically and maitained under standard tissue culture conditions or on collagenI- or tenascin FNIII-coated plates.. Experiment Overall Design: Genetic manipulation (stable transgene expression)
2009-10-03 | E-GEOD-18277 | biostudies-arrayexpress
Project description:RNA-seq to identify targets of integrin alpha2 in breast cancer
| PRJNA549584 | ENA
Project description:To test whether nuclear importin alpha2 can regulate transcription, we sought to examine gene expression changes in cells with nuclear accumulation of importin alpha2 by performing microarray analysis. We designed an experiment in which EGFP-fused full-length importin alpha2 was transfected into HeLa cells. To exclude the possibility that exogenous, full-length importin alpha2 protein enhances nuclear transport of karyophilic proteins such as transcription factors and thereby directly influences gene expression, a mutant of importin alpha2 which is mutated in the C-terminal CAS-binding domain was also transfected, so that it is never recycled to the cytoplasm and shows complete nuclear localization. We undertook to investigate whether there were changes in gene expression common to cells expressing the full-length importin alpha2 and the C-terminal mutant (C-mutant) isoform. EGFP vs. EGFP-importin alpha2 full length, or EGFP vs EGFP-importin alpha2 CAS-binding mutant. One replicate each. EGFP-expressing cells are used as a control.
2010-11-24 | E-GEOD-25303 | biostudies-arrayexpress
Project description:To test whether nuclear importin alpha2 can regulate transcription, we sought to examine gene expression changes in cells with nuclear accumulation of importin alpha2 by performing microarray analysis. We designed an experiment in which EGFP-fused full-length importin alpha2 was transfected into HeLa cells. To exclude the possibility that exogenous, full-length importin alpha2 protein enhances nuclear transport of karyophilic proteins such as transcription factors and thereby directly influences gene expression, a mutant of importin alpha2 which is mutated in the C-terminal CAS-binding domain was also transfected, so that it is never recycled to the cytoplasm and shows complete nuclear localization. We undertook to investigate whether there were changes in gene expression common to cells expressing the full-length importin alpha2 and the C-terminal mutant (C-mutant) isoform.
2010-11-24 | GSE25303 | GEO
Project description:Tumor microenvironment or stroma has the potency to regulate the behavior of malignant cells. Fibroblast-like cells are abundant in tumor stroma and they are also responsible for the synthesis of many extracellular matrix components. Fibroblast–cancer cell interplay can modify the functions of both cell types. We applied mass spectrometry and proteomics to unveil the matrisome in 3D spheroids formed by DU145 prostate cancer cells, PC3 prostate cancer cells or prostate derived fibroblasts. Similarly DU145/fibroblast and PC3/fibroblast co-culture spheroids were also analyzed. Western Blotting and immunofluorescence were used to confirm the presence of specific proteins in spheroids. Cancer dissemination was studied by utilizing "out of spheroids" migration and invasion assays. In the spheroid model cancer cell–fibroblast interplay caused remarkable changes in extracellular matrix and accelerated the invasion of DU145 cells. Fibroblasts produced structural matrix proteins, growth factors and matrix metalloproteinases. In cancer cell/fibroblast co-cultures basement membrane components, including laminins (3, 5, 2, 3), heparan sulphate proteoglycan (HSPG2 gene product), and collagen XVIII accumulated in a prominent manner when compared to spheroids that contained fibroblasts or cancer cells only. Furthermore, collagen XVIII was intensively processed to different endostatin isoforms by cancer cell derived cathepsin L. To sum up, fibroblasts can promote carcinoma cell dissemination by several different mechanisms. Extracellular matrix and basement membrane proteins provide attachment sites for cell locomotion promoting adhesion receptors. Growth factors and metalloproteinases are known to accelerate cell invasion. Additionally, cancer cell–fibroblast interplay generates biologically active fragments of basement membrane proteins, such as endostatin.
2020-05-04 | PXD016882 | Pride
Project description:Previous reports demonstrate that the α2-integrin (α2) mediates pancreatic ductal adenocarcinoma (PDAC) cell interaction with collagens. We found that untransformed pancreatic ductal epithelial cells and well-differentiated PDAC cells use α2 exclusively to adhere and migrate on collagenI. In contrast, poorly-differentiated PT45P1 and MIAPaCa2 cells demonstrate reduced reliance on, or complete loss of α2, respectively. Further, well-differentiated PDAC lines exhibit reduced in vitro invasion compared to poorly-differentiated lines, and α2-blockade suppressed invasion of well-differentiated lines exclusively. Based on these data and the demonstrated role of α2 in maintaining tissue architecture in other organs, we hypothesized that α2 may actually suppress the malignant phenotype in PDAC. Accordingly, stable ectopic expression of α2 in MIAPaCa2 cells (MP2-α2) retarded in vitro invasion. MP2-α2 cells maintained on collagenI were more invasion-retarded than MP2-α2 cells maintained in standard tissue culture, and demonstrated higher α2β1 expression that was reflected in faster and more complete adhesion/migration on collagenI. Affymetrix gene expression profiling of α2-expressing and mock-transfected cells revealed that kallikrein-related peptidases (KLK)-5, 6 and 7 were specifically upregulated by α2. Accordingly, well-differentiated PDAC lines express KLK-5, 6 and 7 and KLK blockade increased invasion as well as collagenI-migration in KLK-positive lines. Importantly, an α2 cytoplasmic deletion mutant promoted KLK expression and retarded invasion, while an α9α2 chimera retarded invasion less efficiently, and did not impact KLK expression. These data demonstrate for the first time that the α2-ectodomain and KLK’s coordinately regulate a less invasive phenotype in PDAC cells.
2009-09-26 | GSE18277 | GEO
Project description:Familial hemiplegic migraine is an episodic neurological disorder characterized by transient sensory and motor symptoms and signs. Mutations of the ion pump alpha2-Na/K ATPase represent a key genetic cause of familial hemiplegic migraine, but the mechanisms by which alpha2-Na/K ATPase mutations lead to the migraine phenotype remain incompletely understood. Here, we unexpectedly find that mice in which alpha2-Na/K ATPase is conditionally deleted in astrocytes display episodic transient motor paralysis. Functional neuroimaging reveals that conditional knockout of alpha2-Na/K ATPase triggers spontaneous cortical spreading depression events that are associated with low voltage activity events upon EEG monitoring, which in turn correlate with transient motor impairment in these mice. Transcriptomic and metabolomic analyses show that loss of alpha2-Na/K ATPase alters metabolic gene expression in astrocytes in vivo with consequent elevation of serine and glycine in the brain. Strikingly, feeding alpha2-Na/K ATPase knockout mice a serine- and glycine-free diet reverses the phenotype of transient motor impairment. Together, our findings define a novel metabolic mechanism regulated by astrocytic alpha2-Na/K ATPase that triggers episodic transient motor paralysis in mice, laying the foundation for potential new treatment strategies for patients with familial hemiplegic migraine.
2020-10-20 | GSE145102 | GEO
Project description:The Illumina Infinium 850k Human DNA methylation BeadChip was used to obtain DNA methylation profiles in DU145 cell samples. Samples included normal, PLK1 knockdown and UHRF1 knockdown DU145 cell samples. Each sample is repeated three times.
2022-10-31 | GSE185400 | GEO