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A Systematic Substrate Screen Links Cyclin D-Dependent Kinases to Senescence Suppression in Cancer Cells through FOXM1

ABSTRACT: Gene expression (mRNA profiling) of U2OS osteosarcoma cells treated with 1 mM of the CDK4/6-specific inhibitor, PD0332991, versus vehicle (DMSO) for 4 hours Here, we monitored the early transcriptional response to acute inhibition of CDK4/6 catalytic activity. As PD0332991 substantially induced senescence in U2OS cells, we performed gene expression arrays from U2OS cells treated with or without PD0332991 for 4 hours. Although this did not result in significant changes in the cell cycle profile, it led to significant changes in global gene expression.

ORGANISM(S): Homo sapiens

PROVIDER: GSE32182 | GEO | 2011/11/18

SECONDARY ACCESSION(S): PRJNA147393

REPOSITORIES: GEO

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A Systematic Substrate Screen Links Cyclin D-Dependent Kinases to Senescence Suppression in Cancer Cells through FOXM1

Project description:Gene expression (mRNA profiling) of U2OS osteosarcoma cells treated with 1 mM of the CDK4/6-specific inhibitor, PD0332991, versus vehicle (DMSO) for 4 hours Here, we monitored the early transcriptional response to acute inhibition of CDK4/6 catalytic activity. As PD0332991 substantially induced senescence in U2OS cells, we performed gene expression arrays from U2OS cells treated with or without PD0332991 for 4 hours. Although this did not result in significant changes in the cell cycle profile, it led to significant changes in global gene expression. 10 total samples were analyzed, i.e. we compared five PD0332991-treated versus five vehicle-treated U2OS cell samples (gene level).

2011-11-18 | E-GEOD-32182 | biostudies-arrayexpress

Ligand-dependent hedgehog signaling maintains an undifferentiated, malignant osteosarcoma phenotype

Project description:To determine the differentially expreesed genes in osteosarcoma cell lines MG63 and U2OS stimulated with hedgehog ligand (SHH) for 24 hours MG63 and U2OS cells were treated with PBS or 1ug/ml of recombinat human sonic hedgehog for 24 hours. RNA was isolated from the cells and RNAseq performed The results present differentially expressed genes when osteosarcoma cell lines are stimulated with SHH

2023-10-16 | GSE192547 | GEO

Gene regulation by m6A demethylase ALKBH5 in U2OS osteosarcoma

Project description:To identify target genes regulated by ALKBH5 in osteosarcoma, we silenced the expression of ALKBH5 in osteosarcoma cell line-U2OS and tested its effect on U2OS transcriptome.

2023-12-31 | GSE171671 | GEO

siRNA profiling of human cell line IMR-32 at time point 0 and transiently transfected with siRNA against GFP, Cyclin D1 and CDK4 at time point 48 hours reveals the undifferentiated phenotype in neuroblastoma depends on Cyclin D1 and CDK4 activity

Project description:Genomic aberrations of Cyclin D1 (CCND1) and CDK4 in neuroblastoma indicate that dysregulation of the G1 entry checkpoint is an important cell cycle aberration in this pediatric tumor. Here we report that analysis of Affymetrix expression data of primary neuroblastic tumors shows an extensive over-expression of Cyclin D1 and CDK4 which correlates with histological subgroups and prognosis respectively. Immunohistochemical analysis demonstrated an over-expression of Cyclin D1 in neuroblasts and a low Cyclin D1 expression in all cell types in ganglioneuroma. This suggests an involvement of G1 regulating genes in neuronal differentiation processes which we further evaluated using RNA interference against Cyclin D1 and its kinase partner CDK4 in several neuroblastoma cell lines. This resulted in pRb pathway inhibition as shown by an almost complete disappearance of CDK4 specific pRb phosphorylation; reduction of E2F transcriptional activity and a decrease of Cyclin A protein levels. The Cyclin D1 and CDK4 knock-down resulted in a significant reduction in cell proliferation, a G1 specific cell cycle arrest and moreover an extensive neuronal differentiation. Affymetrix microarray profiling of siRNA treated cells revealed a shift in expression profile towards a neuronal phenotype. Several new potential downstream players are identified. We conclude that neuroblastoma functionally depend on over-expression of G1 regulating genes to maintain their undifferentiated phenotype. Experiment Overall Design: The Cell line IMR-32 at time point 0 and transiently transfected with siRNA against GFP, Cyclin D1 and CDK4 at time point 48 hours. All experiments are biological triplicates.

2008-10-25 | E-GEOD-8866 | biostudies-arrayexpress

Cdk4 is a critical regulator of age-associated hepatic steatosis.

Project description:Purpose: to identify the impact of cdk4 inhibition on genes and pathways involved in age-associated liver disorders using next generation sequencing on young and control- or PD0332991-treated aged mice. Methods: RNA was isolated from livers of young (2 month old) and old (20-22 month old) C57Bl6 mice. Old mice were either either control-aged mice or PD-0332991 treated aged mice (450mg/dk for 2 weeks). The three conditions were represented in triplicate. Raw reads passing quality filtration were aligned to the mm10 mouse genome using UCSC annotations by pseudoalignment in Kallisto. All reasonably expressed transcripts were submitted for statistical analysis to identify age-altered transcripts corrected by cdk4 inhibition. Results: We mapped a minimum of 20 million reads per sample, which corresponded to a total of 25,240 transcripts. Of these, 12,551 were considered reasonably expressed and were included in analysis comparing control-aged v young and PD0332991-treated-aged v young. Conclusion: Inhibition of cdk4 in ages wildtype mice by cdk4 inhibitor PD-0332991 reduces CEBPa-p300 and reduced aged-related alterations of hepatic structure and function.

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CDK4 phosphorylation status and a linked gene expression profile predict sensitivity to Palbociclib [lines]

Project description:Cyclin D-CDK4/6 are the first CDK complexes to be activated in G1 phase in response to oncogenic pathways. The specific CDK4/6 inhibitor PD0332991 (Palbociclib) was recently approved by the FDA to treat advanced ER+ breast tumors. Unfortunately, reliable predictive tools are lacking to identify potentially responsive or insensitive tumors. We have shown that the activating T172 phosphorylation of CDK4 is the central rate-limiting event that initiates the cell cycle decision and signals the presence of active CDK4. Here, we found that, in breast cancer cell lines, the CDK4 T172 phosphorylation best correlates with the sensitivity to PD0332991. Moreover, the modification profile of CDK4 differs among breast tumors and it associates with their subtypes and risk. A gene expression signature faithfully predicted CDK4 modification profiles in tumors and cell lines. This surrogate biomarker identifies tumors that are unlikely to respond to CDK4/6 inhibitors and could allow extending the indication of the drug to HER-2 positive and some basal-like tumors. Predictive tools to identify breast cancer patients sensitive to Pfizer's new drug targeting CDK4/6 are lacking. We correlated the sensitivity of breast cancer cell lines to the drug to their post-translational modification profile of CDK4 revealed by 2D SDS/poly-acrylamide gel electrophoresis followed by CDK4 immuno-detection and their gene expression profiles. we revealed that breast tumors display similar CDK4 post-translational modification profiles as cell lines and that a gene expression profile can be used to predict the CDK4 post-translational modification profile of a tumor or cell line and thereby its sensitivity to CDK4 inhibitor

2017-01-01 | GSE87006 | GEO

CDK4 phosphorylation status and a linked gene expression profile predict sensitivity to Palbociclib [tumors]

2017-01-01 | GSE87007 | GEO

High-throughput sequencing of OS cells treated with activated compound B5.

Project description:The high-throughput sequencing technology was performed after the treatment of human osteosarcoma cells U2OS and MG63 with the active compound B5 designed and synthesized by ourselves, to explore the expression of genes related to cell adhesion, migration and invasion of human osteosarcoma cells U2OS and MG63 after the treatment of the active compound Changes to explore the effect of active compounds on the motility of osteosarcoma cells.

2020-08-25 | GSE156729 | GEO

Microarray expression profile and functional analysis of circular RNAs in osteosarcoma

Project description:Seven human osteosarcoma cell lines (U2OS, U2OS/MTX300, HOS, MG63, 143B, ZOS, ZOSM) and the human osteoblast hFOB1.19 were included in the study. Microarray based circRNA expression profiles were acquired using the Arraystar Human circRNA Array (8x15K, Arraystar). We identified circRNAs differentially expressed in human osteosarcoma cell lines compared to human osteoblast hFOB1.19 (control).

2017-04-12 | GSE96964 | GEO

RNA sequencing to determine gene expression changes in SQLE knockdown osteosarcoma cells

Project description:In our study, we found that SQLE was critical to osteosarcoma progression. To identify the mechanism of SQLE in osteosarcoma, we conducted the RNA-seq analysis of SQLE knockdown in osteosarcoma cell line U2OS cells.

2024-12-16 | GSE245562 | GEO

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