Relation of exposure to amino acids involved in sarcosine metabolic pathway on behavior of non-tumor and malignant prostatic cell lines

The Prostate ◽  
2016 ◽  
Vol 76 (7) ◽  
pp. 679-690 ◽  
Author(s):  
Zbynek Heger ◽  
Jaromir Gumulec ◽  
Natalia Cernei ◽  
Hana Polanska ◽  
Martina Raudenska ◽  
...  
Keyword(s):  
Amino Acids ◽  
Cell Lines ◽  
Metabolic Pathway ◽  
Prostatic Cell

scholarly journals Diverse metabolic response of cancer cells treated with a 213Bi-anti-EGFR-immunoconjugate

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Benedikt Feuerecker ◽  
Philipp Biechl ◽  
Christof Seidl ◽  
Frank Bruchertseifer ◽  
Alfred Morgenstern ◽  
...  
Keyword(s):  
Amino Acids ◽  
Cancer Cells ◽  
Treatment Response ◽  
Cell Lines ◽  
Egf Receptor ◽  
Metabolic Response ◽  
Gas Chromatography Mass Spectrometry ◽  
Alpha Emitter ◽  
Early Treatment Response ◽  
Bladder Cancer Cells

AbstractEvaluation of treatment response is among the major challenges in modern oncology. We herein used a monoclonal antibody targeting the EGF receptor (EGFR) labelled with the alpha emitter 213Bi (213Bi-anti-EGFR-MAb). EJ28Luc (bladder) and LN18 (glioma) cancer cells, both overexpressing EGFR, were incubated for 3 h with the radioimmunoconjugate. To assess the responses in the core carbon metabolism upon this treatment, these cancer cell lines were subsequently cultivated for 18 h in the presence of [U-13C6]glucose. 13C-enrichment and isotopologue profiles of key amino acids were monitored by gas chromatography–mass spectrometry (GC/MS), in order to monitor the impacts of the radionuclide-treatment upon glucose metabolism. In comparison to untreated controls, treatment of EJ28Luc cells with 213Bi-anti-EGFR-MAb resulted in a significantly decreased incorporation of 13C from [U-13C6]glucose into alanine, aspartate, glutamate, glycine, proline and serine. In sharp contrast, the same amino acids did not display less 13C-enrichments during treatment of the LN18 cells. The data indicate early treatment response of the bladder cancer cells, but not of the glioma cells though cell lines were killed following 213Bi-anti-EGFR-MAb treatment. The pilot study shows that the 13C-labelling approach is a valid tool to assess the responsiveness of cancer cells upon radionuclide-treatment in considerable metabolic detail.

scholarly journals Human T-Cell Leukemia Virus Type 1 Envelope-Mediated Syncytium Formation Can Be Activated in Resistant Mammalian Cell Lines by a Carboxy-Terminal Truncation of the Envelope Cytoplasmic Domain

2003 ◽  
Vol 77 (2) ◽  
pp. 963-969 ◽  
Author(s):  
Felix J. Kim ◽  
Nicolas Manel ◽  
Yvan Boublik ◽  
Jean-Luc Battini ◽  
Marc Sitbon
Keyword(s):  
Amino Acids ◽  
Cell Lines ◽  
Leukemia Virus ◽  
Syncytium Formation ◽  
Cytoplasmic Domain ◽  
Cell Leukemia ◽  
Cell Infection ◽  
Human T Cell ◽  
T Cell Leukemia Virus ◽  
Carboxy Terminal

ABSTRACT Human T-cell leukemia virus (HTLV) envelope (Env) glycoproteins induce fusion, leading to rampant syncytium formation in a broad range of cell lines. Here, we identified murine, hamster, canine, and porcine cell lines that are resistant to HTLV-1 Env-induced syncytium formation. This resistance was not due to the absence of functional receptors for HTLV Env, as these cells were susceptible to infection with HTLV Env-pseudotyped virions. As murine leukemia virus (MLV) Env and HTLV Env present close structural homologies (F. J. Kim, I. Seiliez, C. Denesvre, D. Lavillette, F. L. Cosset, and M. Sitbon, J. Biol. Chem. 275:23417-23420, 2000), and because activation of syncytium formation by MLV Env generally requires cleavage of the R peptide in the cytoplasmic domain of the Env transmembrane (TM) component, we assessed whether truncation of the cytoplasmic domain of HTLV Env would alleviate this resistance. Indeed, in all resistant cell lines, truncation of the last 8 amino acids of the HTLV Env cytoplasmic domain (HdC8) was sufficient to overcome resistance to HTLV Env-induced syncytium formation. Furthermore, HdC8-mediated cell-to-cell infection titers varied according to the target cell lines and could be significantly higher than that observed with HTLV Env on HeLa cells. These data indicate that a determinant located within the 8 carboxy-terminal cytoplasmic amino acids of TM plays a distinct role in HTLV Env-mediated cell-to-cell infection and syncytium formation.

scholarly journals Validation of commercially available sphingosine kinase 2 antibodies for use in immunoblotting, immunoprecipitation and immunofluorescence

F1000Research ◽  
2016 ◽  
Vol 5 ◽  
pp. 2825 ◽  
Author(s):  
Heidi A. Neubauer ◽  
Stuart M. Pitson
Keyword(s):  
Amino Acids ◽  
Cell Lines ◽  
Polyclonal Antibody ◽  
Human Cell ◽  
Sphingosine Kinase ◽  
Immunofluorescence Staining ◽  
Human Cell Lines ◽  
Sphingosine Kinase 2 ◽  
Hela Cell Lines ◽  
Rabbit Polyclonal Antibody

Sphingosine kinase 2 (SK2) is a ubiquitously expressed lipid kinase that has important, albeit complex and poorly understood, roles in regulating cell survival and cell death. In addition to being able to promote cell cycle arrest and apoptosis under certain conditions, it has recently been shown that SK2 can promote neoplastic transformation and tumorigenesis in vivo. Therefore, well validated and reliable tools are required to study and better understand the true functions of SK2. Here, we compare two commercially available SK2 antibodies: a rabbit polyclonal antibody from Proteintech that recognizes amino acids 266-618 of human SK2a, and a rabbit polyclonal antibody from ECM Biosciences that recognizes amino acids 36-52 of human SK2a. We examine the performance of these antibodies for use in immunoblotting, immunoprecipitation and immunofluorescence staining of endogenous SK2, using human HEK293 and HeLa cell lines, as well as mouse embryonic fibroblasts (MEFs). Furthermore, we assess the specificity of these antibodies to the target protein through the use of siRNA-mediated SK2 knockdown and SK2 knockout (Sphk2-/-) MEFs. Our results demonstrate that the Proteintech anti-SK2 antibody reproducibly displayed superior sensitivity and selectivity towards SK2 in immunoblot analyses, while the ECM Biosciences anti-SK2 antibody was reproducibly superior for SK2 immunoprecipitation and detection by immunofluorescence staining. Notably, both antibodies produced non-specific bands and staining in the MEFs, which was not observed with the human cell lines. Therefore, we conclude that the Proteintech SK2 antibody is a valuable reagent for use in immunoblot analyses, and the ECM Biosciences SK2 antibody is a useful tool for SK2 immunoprecipitation and immunofluorescence staining, at least in the human cell lines employed in this study.

Characterization and expression of the human rhoH12 gene product

1989 ◽  
Vol 9 (5) ◽  
pp. 2058-2066
Author(s):  
H Avraham ◽  
R A Weinberg
Keyword(s):  
Amino Acids ◽  
Cell Lines ◽  
Soft Agar ◽  
Cdna Clones ◽  
Focus Formation ◽  
Reductase Gene ◽  
Evolutionarily Conserved ◽  
Normal Human ◽  
Nih 3T3 ◽  
P21 Proteins

The rho genes constitute an evolutionarily conserved family having significant homology to the ras oncogene family. These genes have been found in Saccharomyces cerevisiae, Drosophila melanogaster, rat, and human; their 21,000-dalton products show strong conservation of structure. In humans, three classes of rho cDNA clones have been identified which differ by virtue of the presence of variable C-terminal domains: rhoH12, rhoH6, and rhoH9. The predicted 193 amino acids of human rhoH12 protein show 88% similarity with those of the human rhoH6 clone, 96.8% similarity with those of the Aplysia rho product, and 81.8% similarity with those of the yeast RHO1 protein. Rat-1 and NIH 3T3 mouse fibroblasts were transfected with clones containing the normal human rhoH12 allele as well as the variants encoding valine in place of the glycine and leucine in place of the glutamine normally found at residues 14 and 64, respectively. These replacements mirror the changes responsible for oncogenic activation of the related ras-encoded p21 proteins. These mutant rhoH12 clone alleles did not cause focus formation in monolayers or growth in soft agar. However, amplification of normal rhoH12 via cotransfection with a dihydrofolate reductase gene resulted in colonies that displayed reduced dependence on serum for growth, grew to higher saturation densities, and were tumorigenic when inoculated into nude mice. Normal p21rho protein was detected in the transfected cell lines as well as in normal cell lines by Western immunoblot and immunoprecipitation analysis with rabbit antibodies raised against the peptide corresponding to amino acids 122 to 135.

The Effect of Cell Population Density on Nutrient Uptake and Cell Metabolism: a Comparative Study of Human Diploid and Heteroploid Cell Lines

1972 ◽  
Vol 10 (2) ◽  
pp. 515-524
Author(s):  
J. B. GRIFFITHS
Keyword(s):  
Amino Acids ◽  
Protein Synthesis ◽  
Amino Acid ◽  
Dna Synthesis ◽  
Nutrient Uptake ◽  
Cell Lines ◽  
Acid Concentration ◽  
Contact Inhibition ◽  
Inhibition Of Growth ◽  
Extracellular Amino Acid

The possibility that contact inhibition of growth in cultures of human diploid cells is influenced by the effects of cell crowding on nutrient uptake by the cells was investigated. Two human lung cell lines were compared, the diploid line MRC-5 and the heteroploid line L-132. In pre-confluent cultures the ability of these 2 cell types to accumulate amino acids was very similar. Post-confluent L-132 cells showed very little change from the pre-confluent cultures but the ability of MRC-5 cells in post-confluent cultures was greatly reduced. The intracellular concentrations of various amino acids necessary to achieve the maximum rate of protein synthesis were found. These values were identical for sparse and crowded cultures but due to the reduced uptake ability of crowded MRC-5 cells a far higher external amino acid concentration was required in post-confluent cultures. This meant that although amino acids did not become growth-limiting until over 80% utilized in pre-confluent cultures, in post-confluent cultures they became growth-limiting when only 50% utilized. Although protein synthesis was significantly affected by extracellular amino acid concentration and cell crowding, thus contributing towards the effect of contact inhibition of growth, DNA synthesis was shown to be the major metabolic function in contact inhibition. Increased cell density had a very inhibitory effect on DNA synthesis in MRC-5 cultures, but not in L-132 cultures, and this was unaffected by extracellular amino acid and glucose concentration.

scholarly journals Prognostic Significance of Serum Free Amino Acids in Head and Neck Cancers

Cells ◽  
2019 ◽  
Vol 8 (5) ◽  
pp. 428 ◽  
Author(s):  
Vit Vsiansky ◽  
Marketa Svobodova ◽  
Jaromir Gumulec ◽  
Natalia Cernei ◽  
Dagmar Sterbova ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Regression Model ◽  
Head And Neck ◽  
Cell Lines ◽  
Proportional Hazards ◽  
Cox Proportional Hazards ◽  
Transformed Cells ◽  
Cox Proportional Hazards Regression ◽  
Proportional Hazards Regression

Despite distinctive advances in the field of head and neck squamous cell cancer (HNSCC) biomarker discovery, the spectrum of clinically useful prognostic serum biomarkers is limited. As metabolic activities in highly proliferative transformed cells are fundamentally different from those in non-transformed cells, specific shifts in concentration of different metabolites may serve as diagnostic or prognostic markers. Blood amino acids have been identified as promising biomarkers in different cancers before, but little is known about this field in HNSCC. Blood amino acid profiles of 140 HNSCC patients were examined using high-performance liquid chromatography. Cox proportional hazards regression model was used to assess the prognostic value of amino acid concentrations in serum. Colony forming assay was used to identify the effect of amino acids that were significant in Cox proportional hazards regression models on colony forming ability of FaDu and Detroit 562 cell lines. In the multivariable Cox regression model for overall survival (OS), palliative treatment was associated with an unfavourable prognosis while high serum levels of methionine have had a positive prognostic impact. In the relapse-free survival (RFS) multivariable model, methionine was similarly identified as a positive prognostic factor, along with tumor localization in the oropharynx. Oral cavity localization and primary radio(chemo)therapy treatment strategy have been linked to poorer RFS. 1mM serine was shown to support the forming of colonies in both tested HNSCC cell lines. Effect of methionine was exactly the opposite.

Fatty acid recycling in adipocytes: a role for glyceroneogenesis and phosphoenolpyruvate carboxykinase

2003 ◽  
Vol 31 (6) ◽  
pp. 1125-1129 ◽  
Author(s):  
C. Forest ◽  
J. Tordjman ◽  
M. Glorian ◽  
E. Duplus ◽  
G. Chauvet ◽  
...  
Keyword(s):  
Type 2 Diabetes ◽  
Amino Acids ◽  
Adipose Tissue ◽  
Fatty Acid ◽  
White Adipose Tissue ◽  
Metabolic Pathway ◽  
Phosphoenolpyruvate Carboxykinase ◽  
Important Pathway ◽  
Low Activity

FA (fatty acid) recycling in adipose tissue appears to be an important pathway for regulating FA release into the blood during fasting. Re-esterification requires G3P (glycerol 3-phosphate), which cannot be synthesized from glucose because glycolysis is much reduced under such circumstances. In addition, G3P can scarcely originate from glycerol since glycerol kinase has a very low activity in white adipose tissue. It was shown about 35 years ago that a metabolic pathway named glyceroneogenesis, which allows G3P synthesis from non-carbohydrate precursors like pyruvate, lactate or amino acids, is activated during fasting. The major enzyme in this pathway was shown to be PEPCK-C [cytosolic phosphoenolpyruvate carboxykinase (GTP); EC 4.1.1.32]. The present review analyses the mechanisms by which a series of hormones and nutrients affect PEPCK-C gene transcription and glyceroneogenesis and describes evidence for dysregulation of this pathway in type 2 diabetes.

Small interference RNA targeting heat-shock protein 27 inhibits the growth of prostatic cell lines and induces apoptosis via caspase-3 activation in vitro

2006 ◽  
Vol 98 (5) ◽  
pp. 1082-1089 ◽  
Author(s):  
Palma Rocchi ◽  
Paul Jugpal ◽  
Alan So ◽  
Shannon Sinneman ◽  
Susan Ettinger ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Cell Lines ◽  
Caspase 3 ◽  
Heat Shock Protein 27 ◽  
Small Interference Rna ◽  
Rna Targeting ◽  
Interference Rna ◽  
Prostatic Cell

scholarly journals Metabolic Pathway Confirmation and Discovery Through 13C-labeling of Proteinogenic Amino Acids

10.3791/3583 ◽  
2012 ◽  
Author(s):  
Le You ◽  
Lawrence Page ◽  
Xueyang Feng ◽  
Bert Berla ◽  
Himadri B. Pakrasi ◽  
...  
Keyword(s):  
Amino Acids ◽  
Metabolic Pathway

Quantification of integrin subunits on human prostatic cell lines—Comparison of nontumorigenic and tumorigenic lines

The Prostate ◽  
1997 ◽  
Vol 31 (1) ◽  
pp. 1-8 ◽  
Author(s):  
Patricia L. Haywood-Reid ◽  
David R. Zipf ◽  
Wayne R. Springer
Keyword(s):  
Cell Lines ◽  
Prostatic Cell
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