Knockout of the caspase 8‐associated protein 2 gene improves recombinant protein expression in HEK293 cells through up‐regulation of the cyclin‐dependent kinase inhibitor 2A gene

2020 ◽  
Vol 118 (1) ◽  
pp. 186-198
Author(s):  
Laura Abaandou ◽  
Ashish K. Sharma ◽  
Joseph Shiloach
Keyword(s):  
Recombinant Protein ◽  
Protein Expression ◽  
Kinase Inhibitor ◽  
Recombinant Protein Expression ◽  
Hek293 Cells ◽  
Caspase 8 ◽  
Cyclin Dependent Kinase ◽  
Cyclin Dependent Kinase Inhibitor

scholarly journals Frequency of polymorphisms and protein expression of cyclin-dependent kinase inhibitor 1A (CDKN1A) in central nervous system tumors

2009 ◽  
Vol 127 (5) ◽  
pp. 288-294 ◽  
Author(s):  
Mev Dominguez Valentin ◽  
Renata Canalle ◽  
Rosane de Paula Queiroz ◽  
Luiz Gonzaga Tone
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Protein Expression ◽  
Kinase Inhibitor ◽  
Expression Patterns ◽  
Cns Tumors ◽  
Control Group ◽  
Cyclin Dependent Kinase ◽  
Cross Sectional ◽  
Cyclin Dependent Kinase Inhibitor

CONTEXT AND OBJECTIVE: Genetic investigation of central nervous system (CNS) tumors provides valuable information about the genes regulating proliferation, differentiation, angiogenesis, migration and apoptosis in the CNS. The aim of our study was to determine the prevalence of genetic polymorphisms (codon 31 and 3' untranslated region, 3'UTR) and protein expression of the cyclin-dependent kinase inhibitor 1A (CDKN1A) gene in patients with and without CNS tumors. DESIGN AND SETTING: Analytical cross-sectional study with a control group, at the Molecular Biology Laboratory, Pediatric Oncology Department, Hospital das Clínicas de Ribeirão Preto. METHODS: 41 patients with CNS tumors and a control group of 161 subjects without cancer and paires for sex, age and ethnicity were genotyped using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Protein analysis was performed on 36 patients with CNS tumors, using the Western Blotting technique. RESULTS: The frequencies of the heterozygote (Ser/Arg) and polymorphic homozygote (Arg/Arg) genotypes of codon 31 in the control subjects were 28.0% and 1.2%, respectively. However, the 3'UTR site presented frequencies of 24.2% (C/T) and 0.6% (T/T). These frequencies were not statistically different (P > 0.05) from those seen in the patients with CNS tumors (19.4% and 0.0%, codon 31; 15.8% and 2.6%, 3'UTR site). Regarding the protein expression in ependymomas, 66.67% did not express the protein CDKN1A. The results for medulloblastomas and astrocytomas were similar: neither of them expressed the protein (57.14% and 61.54%, respectively). CONCLUSION: No significant differences in protein expression patterns or polymorphisms of CDKN1A in relation to the three types of CNS tumors were observed among Brazilian subjects.

scholarly journals MIB-1 (KI-67) Proliferation Index and Cyclin-Dependent Kinase Inhibitor p27Kip1 Protein Expression in Nephroblastoma

2004 ◽  
Vol 10 (2) ◽  
pp. 591-597 ◽  
Author(s):  
Mazen A. Ghanem ◽  
Theo H. Van der Kwast ◽  
Mondastri K. Sudaryo ◽  
Rejiv B. Mathoera ◽  
Marry M. van den Heuvel ◽  
...  
Keyword(s):  
Protein Expression ◽  
Kinase Inhibitor ◽  
Proliferation Index ◽  
Cyclin Dependent Kinase ◽  
Ki 67 ◽  
P27kip1 Protein ◽  
Cyclin Dependent Kinase Inhibitor ◽  
Mib 1

Application of Proteomics in the Search for Novel Proteins Associated with the Anti-cancer Effect of the Synthetic Cyclin-dependent Kinases Inhibitor, Bohemine

2002 ◽  
Vol 1 (4) ◽  
pp. 247-256 ◽  
Author(s):  
Hana Kovarova ◽  
Petr Halada ◽  
Petr Man ◽  
Petr Dzubak ◽  
Marian Hajduch
Keyword(s):  
Mass Spectrometry ◽  
Cell Cycle ◽  
Protein Expression ◽  
Cell Line ◽  
Metabolic Pathways ◽  
Kinase Inhibitor ◽  
A549 Cells ◽  
Cyclin Dependent Kinase ◽  
Cyclin Dependent Kinase Inhibitor ◽  
Anti Cancer

The purpose of this study was to use the proteomics approach, which is based on high resolution two-dimensional electrophoresis coupled with multivariate correspondence analysis and mass spectrometry, to classify objectively the biochemical basis of the anti-cancer activity of the synthetic cyclin-dependent kinase inhibitor, bohemine (BOH). The changes in the cell cycle and corresponding protein composition of the A549 human lung adenocarcinoma cell line after treatment with BOH were evaluated and proteins differentially expressed in the BOH treated A549 cells, compared to the untreated A549 counterparts, were selected. Thirteen of these candidate proteins associated with the drug effects in vitro were identified by mass spectrometry. Many of these proteins fall into one of three functional categories: i) metabolic pathways (glycolysis, nucleic acid synthesis and NADPH production), ii) stress response and protein folding, and iii) cytoskeleton and exocytosis. Changes in protein expression patterns corresponded to a higher resistance of A549 lung carcinoma cells to BOH when compared to the CEM leukaemia cell line. These protein changes reflect a fine balance of the resistant versus the susceptible phenotype in response to the drug. Since BOH is a selective cyclin-dependent kinase inhibitor, changes in the protein expression pattern can be more generally associated with cell cycle regulation as evidenced by inhibition of cell cycling in A549 cells. Our conclusions further underline the importance of cell cycle control in both the cellular signalling and metabolic pathways.

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