Dataset Information

CDK1 is up-regulated by temozolomide in an NF-?B dependent manner in glioblastoma.

ABSTRACT: The alkylating agent, temozolomide (TMZ), is the most commonly used chemotherapeutic for the treatment of glioblastoma (GBM). The anti-glioma effect of TMZ involves a complex response that includes G2-M cell cycle arrest and cyclin-dependent kinase 1 (CDK1) activation. While CDK1 phosphorylation is a well-described consequence of TMZ treatment, we find that TMZ also robustly induces CDK1 expression. Analysis of this pathway demonstrates that CDK1 is regulated by NF-?B via a putative ?B-site in its proximal promoter. CDK1 was induced in a manner dependent on mature p50 and the atypical inhibitor ?B protein, BCL-3. Treatment with TMZ induced binding of NF-?B to the ?B-site as assessed by gel shift analysis and chromatin immunoprecipitation. Examination of a CDK1 promoter-reporter demonstrated the functional relevance of the ?B-site and underlined the requirement of p50 and BCL-3 for activation. Targeted knockdown of CDK1 or chemical inhibition with the selective CDK1 inhibitor, RO-3306, potentiated the cytotoxic effect of TMZ. These results identify CDK1 as an NF-?B target gene regulated by p50 and BCL-3 and suggest that targeting CDK1 may be a strategy to improve the efficacy of TMZ against GBM.

SUBMITTER: Voce DJ

PROVIDER: S-EPMC7952566 | biostudies-literature | 2021 Mar

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

CDK1 is up-regulated by temozolomide in an NF-κB dependent manner in glioblastoma.

Voce David J DJ Bernal Giovanna M GM Cahill Kirk E KE Wu Longtao L Mansour Nassir N Crawley Clayton D CD Campbell Paige-Ashley S PS Arina Ainhoa A Weichselbaum Ralph R RR Yamini Bakhtiar B

Scientific reports 20210311 1

The alkylating agent, temozolomide (TMZ), is the most commonly used chemotherapeutic for the treatment of glioblastoma (GBM). The anti-glioma effect of TMZ involves a complex response that includes G2-M cell cycle arrest and cyclin-dependent kinase 1 (CDK1) activation. While CDK1 phosphorylation is a well-described consequence of TMZ treatment, we find that TMZ also robustly induces CDK1 expression. Analysis of this pathway demonstrates that CDK1 is regulated by NF-κB via a putative κB-site in i ...[more]

PMID: 33707466

Similar Datasets

Project description:BackgroundMost DNA-damaging chemotherapeutic agents activate the transcription factor nuclear factor ?B (NF-?B). However, NF-?B activation can either protect from or contribute to the growth suppressive effects of the agent. We previously showed that the DNA-methylating drug temozolomide (TMZ) activates AKT, a positive modulator of NF-?B, in a mismatch repair (MMR) system-dependent manner. Here we investigated whether NF-?B is activated by TMZ and whether AKT is involved in this molecular event. We also evaluated the functional consequence of inhibiting NF-?B on tumor cell response to TMZ.MethodsAKT phosphorylation, NF-?B transcriptional activity, I?B-? degradation, NF-?B2/p52 generation, and RelA and NF-?B2/p52 nuclear translocation were investigated in TMZ-treated MMR-deficient (HCT116, 293TL?-) and/or MMR-proficient (HCT116/3-6, 293TL?+, M10) cells. AKT involvement in TMZ-induced activation of NF-?B was addressed in HCT116/3-6 and M10 cells transiently transfected with AKT1-targeting siRNA or using the isogenic MMR-proficient cell lines pUSE2 and KD12, expressing wild type or kinase-dead mutant AKT1. The effects of inhibiting NF-?B on sensitivity to TMZ were investigated in HCT116/3-6 and M10 cells using the NF-?B inhibitor NEMO-binding domain (NBD) peptide or an anti-RelA siRNA.ResultsTMZ enhanced NF-?B transcriptional activity, activated AKT, induced I?B-? degradation and RelA nuclear translocation in HCT116/3-6 and M10 but not in HCT116 cells. In M10 cells, TMZ promoted NF-?B2/p52 generation and nuclear translocation and enhanced the secretion of IL-8 and MCP-1. TMZ induced RelA nuclear translocation also in 293TL?+ but not in 293TL?- cells. AKT1 silencing inhibited TMZ-induced I?B-? degradation and NF-?B2/p52 generation. Up-regulation of NF-?B transcriptional activity and nuclear translocation of RelA and NF-?B2/p52 in response to TMZ were impaired in KD12 cells. RelA silencing in HCT116/3-6 and M10 cells increased TMZ-induced growth suppression. In M10 cells NBD peptide reduced basal NF-?B activity, abrogated TMZ-induced up-regulation of NF-?B activity and increased sensitivity to TMZ. In HCT116/3-6 cells, the combined treatment with NBD peptide and TMZ produced additive growth inhibitory effects.ConclusionNF-?B is activated in response to TMZ in a MMR- and AKT-dependent manner and confers protection against drug-induced cell growth inhibition. Our findings suggest that a clinical benefit could be obtained by combining TMZ with NF-?B inhibitors.

Dataset Information

CDK1 is up-regulated by temozolomide in an NF-?B dependent manner in glioblastoma.

Publications

CDK1 is up-regulated by temozolomide in an NF-κB dependent manner in glioblastoma.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets