Metabolic Fingerprinting of Murine L929 Fibroblasts as a Cell-Based Tumour Suppressor Model System for Methionine Restriction

Since Otto Warburg reported in 1924 that cancer cells address their increased energy requirement through a massive intake of glucose, the cellular energy level has offered a therapeutic anticancer strategy. Methionine restriction (MetR) is one of the most effective approaches for inducing low-energy metabolism (LEM) due to the central position in metabolism of this amino acid. However, no simple in vitro system for the rapid analysis of MetR is currently available, and this study establishes the murine cell line L929 as such a model system. L929 cells react rapidly and efficiently to MetR, and the analysis of more than 150 different metabolites belonging to different classes (amino acids, urea and tricarboxylic acid cycle (TCA) cycles, carbohydrates, etc.) by liquid chromatography/mass spectrometry (LC/MS) defines a metabolic fingerprint and enables the identification of specific metabolites representing normal or MetR conditions. The system facilitates the rapid and efficient testing of potential cancer therapeutic metabolic targets. To date, MS studies of MetR have been performed using organisms and yeast, and the current LC/MS analysis of the intra- and extracellular metabolites in the murine cell line L929 over a period of 5 days thus provides new insights into the effects of MetR at the cellular metabolic level.

Download Full-text

Remodeling of the fibroblast cytoskeletal architecture during the replication cycle of Ectromelia virus: A morphological in vitro study in a murine cell line

Cytoskeleton ◽

10.1002/cm.21308 ◽

2016 ◽

Vol 73 (8) ◽

pp. 396-417 ◽

Cited By ~ 6

Author(s):

Lidia Szulc-Dabrowska ◽

Karolina P. Gregorczyk ◽

Justyna Struzik ◽

Anna Boratynska-Jasinska ◽

Joanna Szczepanowska ◽

...

Keyword(s):

Cell Line ◽

In Vitro Study ◽

Replication Cycle ◽

Vitro Study ◽

Ectromelia Virus ◽

Murine Cell ◽

Murine Cell Line

Download Full-text

Transcriptional regulation of two myeloid-specific genes, myeloperoxidase and lactoferrin, during differentiation of the murine cell line 32D C13

Blood ◽

10.1182/blood.v78.9.2426.bloodjournal7892426 ◽

1991 ◽

Vol 78 (9) ◽

pp. 2426-2432 ◽

Cited By ~ 1

Author(s):

AD Friedman ◽

BL Krieder ◽

D Venturelli ◽

G Rovera

Keyword(s):

Transcriptional Regulation ◽

Cell Line ◽

Northern Blot ◽

Transcription Initiation ◽

Cell System ◽

Myeloid Differentiation ◽

Murine Cell ◽

Murine Cell Line ◽

Extension Analysis

The transcriptional regulation of myeloperoxidase (MPO) and lactoferrin (LF) was examined during terminal myeloid differentiation of the murine cell line 32D C13. The rates of transcription initiation for MPO and LF, determined by an in vitro nuclear run-on assay, increased approximately ninefold. The accumulation of MPO mRNA in 32D C13 cells, determined by Northern blot analysis, correlated temporally with the observed increase in MPO transcription initiation. On the other hand, accumulation of LF mRNA lagged behind the observed increase in LF transcription initiation. In mouse L cells, the LF gene was transcribed more frequently than the MPO gene, though neither mRNA accumulated. Finally, murine MPO transcription is shown, by Northern blot and primer extension analysis, to initiate at multiple sites. These results indicate that whereas transcription induction may largely account for the accumulation of MPO mRNA during terminal myeloid differentiation, both transcriptional and posttranscriptional mechanisms operate to allow accumulation of LF mRNA. The 32D C13 cell system will be a useful model for elucidating these mechanisms.

Download Full-text

Transcriptional regulation of two myeloid-specific genes, myeloperoxidase and lactoferrin, during differentiation of the murine cell line 32D C13

Blood ◽

10.1182/blood.v78.9.2426.2426 ◽

1991 ◽

Vol 78 (9) ◽

pp. 2426-2432 ◽

Cited By ~ 25

Author(s):

AD Friedman ◽

BL Krieder ◽

D Venturelli ◽

G Rovera

Keyword(s):

Transcriptional Regulation ◽

Cell Line ◽

Northern Blot ◽

Transcription Initiation ◽

Cell System ◽

Myeloid Differentiation ◽

Murine Cell ◽

Murine Cell Line ◽

Extension Analysis

Abstract The transcriptional regulation of myeloperoxidase (MPO) and lactoferrin (LF) was examined during terminal myeloid differentiation of the murine cell line 32D C13. The rates of transcription initiation for MPO and LF, determined by an in vitro nuclear run-on assay, increased approximately ninefold. The accumulation of MPO mRNA in 32D C13 cells, determined by Northern blot analysis, correlated temporally with the observed increase in MPO transcription initiation. On the other hand, accumulation of LF mRNA lagged behind the observed increase in LF transcription initiation. In mouse L cells, the LF gene was transcribed more frequently than the MPO gene, though neither mRNA accumulated. Finally, murine MPO transcription is shown, by Northern blot and primer extension analysis, to initiate at multiple sites. These results indicate that whereas transcription induction may largely account for the accumulation of MPO mRNA during terminal myeloid differentiation, both transcriptional and posttranscriptional mechanisms operate to allow accumulation of LF mRNA. The 32D C13 cell system will be a useful model for elucidating these mechanisms.

Download Full-text