CDK8 knockdown in HT-29 cells and CDK8 & MED12 knockdown in R1 cells
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus Homo sapiens
PROVIDER: GSE30817 | GEO | 2012/03/05
SECONDARY ACCESSION(S): PRJNA144197
REPOSITORIES: GEO
Similar Datasets
Project description:This SuperSeries is composed of the following subset Series: GSE30815: CDK8 knockdown in HT-29 human colon cancer cells GSE30816: CDK8 and MED12 knockdown in R1 mouse ES cells Refer to individual Series
2012-03-05 | E-GEOD-30817 | biostudies-arrayexpress
Project description:Transcriptional profiling of R1 mouse ES cells infected with non-targeting control (NTC) shRNA and two different shRNA sequences against Cdk8 (shCdk8-1 and shCdk-2) and Med12 (shMed12-1 and shMed12-2). Multi-condition experiment comparing shNTC versus shCdk8-1 and shCdk8-2 after 8 days (3 biological replicates each) and shNTC versus shCdk8-1, shCdk8-2, shMed12-1, and shMed12-2 after 13 days (3 biological replicates each); in total 24 samples profiled.
2012-03-05 | E-GEOD-30816 | biostudies-arrayexpress
Project description:Transcriptional profiling of R1 mouse ES cells infected with non-targeting control (NTC) shRNA and two different shRNA sequences against Cdk8 (shCdk8-1 and shCdk-2) and Med12 (shMed12-1 and shMed12-2).
2012-03-05 | GSE30816 | GEO
Project description:Transcriptional profiling of HT-29 human colon cancer cells transfected with non-targeting control (NTC) siRNA and two different siRNA sequences against CDK8 (siCDK8-1 and siCDK8-2).
2012-03-05 | GSE30815 | GEO
Project description:Transcriptional profiling of HT-29 human colon cancer cells transfected with non-targeting control (NTC) siRNA and two different siRNA sequences against CDK8 (siCDK8-1 and siCDK8-2). Three condition experiment comparing siNTC versus siCDK8-1 and siCDK8-2 after 3 days (3 biological replicates each); in total 9 samples profiled.
2012-03-05 | E-GEOD-30815 | biostudies-arrayexpress
Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series
2016-08-25 | E-GEOD-76055 | biostudies-arrayexpress
Project description:The Mediator kinase module regulates eukaryotic transcription by phosphorylating transcription-related targets and by modulating the association of Mediator and RNA polymerase II. The activity of its catalytic core, cyclin-dependent kinase 8 (CDK8), is controlled by Cyclin C and regulatory subunit MED12, with its deregulation contributing to numerous malignancies. Here, we combine in vitro biochemistry, cross-linking coupled to mass spectrometry, and in vivo studies to describe the binding location of the N-terminal segment of MED12 on the CDK8/Cyclin C complex and to gain mechanistic insights into the activation of CDK8 by MED12. Our data demonstrate that the N-terminal portion of MED12 wraps around CDK8, whereby it positions an "activation helix" close to the T-loop of CDK8 for its activation. Intriguingly, mutations in the activation helix that are frequently found in cancers do not diminish the affinity of MED12 for CDK8, yet likely alter the exact positioning of the activation helix. Furthermore, we find the transcriptome-wide gene-expression changes in human cells that result from a mutation in the MED12 activation helix to correlate with deregulated genes in breast and colon cancer. Finally, functional assays in the presence of kinase inhibitors reveal that binding of MED12 remodels the active site of CDK8 and thereby precludes the inhibition of ternary CDK8 complexes by type II kinase inhibitors. Taken together, our results not only allow us to propose a revised model of how CDK8 activity is regulated by MED12, but also offer a path forward in developing small molecules that target CDK8 in its MED12-bound form.
| S-EPMC7022176 | biostudies-literature
Project description:Kallikrein‑related peptidase 12 (KLK12) is overexpressed in cancer tissues including gastric, breast and prostate cancer. However, the role of KLK12 in colorectal cancer is not fully understood. In the present study, the level of KLK12 was determined by performing reverse transcription‑polymerase chain reaction (RT‑qPCR) in colorectal cancer tissues and cell lines. Lipofectamine® 2000 was used to transfect HT‑29 cells to overexpress and knockdown KLK12. Cell viability, migration, invasion and apoptosis were detected by MTT, wound healing, Transwell and flow cytometry assays, respectively. The mRNA and protein expression levels of EMT‑associated proteins, apoptosis‑associated proteins, phosphorylated adenosine monophosphate‑activated protein kinase (p‑AMPK) and phosphorylated mammalian target of rapamycin (p‑mTOR) were determined by RT‑qPCR and western blot analysis. It was identified that the KLK12 mRNA levels were increased significantly in colorectal cancer tissues and cell lines. KLK12 small interfering RNA inhibited cell viability, migration and invasion. Furthermore, epithelial‑mesenchymal transition (EMT)‑associated proteins were altered by siKLK12. Cell apoptosis was induced by KLK12 downregulation, which was demonstrated by the changes in apoptosis‑associated proteins; however, KLK12 overexpression produced the opposite effect. SiKLK12 enhanced the expression of p‑AMPK and suppressed the expression of p‑mTOR, while KLK12 overexpression had the opposite effect. Promotion of KLK12 overexpression‑induced cell viability was reversed by 5‑aminoimidazole‑4‑carboxamide ribonucleotide, an activator of the AMPK signaling pathway, and rapamycin, a specific inhibitor of the mTOR signaling pathway. Taken together, the results of the present study indicated that KLK12 was overexpressed in colorectal cancer and may regulate cell behavior, potentially via the AMPK and mTOR pathways.
| S-EPMC6777684 | biostudies-literature
Project description:This SuperSeries is composed of the SubSeries listed below.
2016-08-25 | GSE76055 | GEO
Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series
2014-03-29 | E-GEOD-40987 | biostudies-arrayexpress