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

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.

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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.

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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

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

International Journal of Molecular Sciences ◽

10.3390/ijms22063039 ◽

2021 ◽

Vol 22 (6) ◽

pp. 3039

Author(s):

Werner Schmitz ◽

Corinna Koderer ◽

Mohamed El-Mesery ◽

Sebastian Gubik ◽

Rene Sampers ◽

...

Keyword(s):

Cell Line ◽

Tricarboxylic Acid Cycle ◽

Energy Requirement ◽

Model System ◽

Central Position ◽

Murine Cell ◽

Murine Cell Line ◽

Methionine Restriction ◽

Metabolic Fingerprint

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.

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Ultrastructure of Choriocarcinoma in Vitro

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010006372x ◽

1969 ◽

Vol 27 ◽

pp. 362-363

Author(s):

John C. Garancis ◽

R. A. Pattillo

Keyword(s):

Cell Line ◽

Physiological Function ◽

Cell System ◽

Bewo Cell ◽

Bewo Cells ◽

Gestational Choriocarcinoma ◽

Bewo Cell Line

Growth of cell system (BeWo-cell line) derived from human gestational choriocarcinoma has been established and continuously maintained in-vitro. Furthermore, it is evident from the previous studies that this cell line has retained the physiological function of the placental trophoblasts, namely the synthesis of human chorionic gonadotrophil(HCG).The BeWo cells were relatively small and possessed single nuclei, thus indicating that this cell line consists exclusively of cytotrophoblasts. In some instances cells appeared widely separated and their lateral surfaces were provided with numerous microvilli (Fig.1).

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Replication of the dihydrofolate reductase genes on double minute chromosomes in a murine cell line

Experimental Cell Research ◽

10.1016/0014-4827(90)90293-j ◽

1990 ◽

Vol 188 (1) ◽

pp. 164-168 ◽

Cited By ~ 3

Author(s):

Thea D. Tlsty ◽

Paula Adams

Keyword(s):

Cell Line ◽

Dihydrofolate Reductase ◽

Murine Cell ◽

Murine Cell Line ◽

Double Minute ◽

Double Minute Chromosomes

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Whole-genome fingerprint of the DNA methylome during chemically induced differentiation of the human AML cell line HL-60/S4

10.1101/608695 ◽

2019 ◽

Author(s):

Enoch Boasiako Antwi ◽

Ada Olins ◽

Vladimir B Teif ◽

Matthias Bieg ◽

Tobias Bauer ◽

...

Keyword(s):

Cell Differentiation ◽

Cell Line ◽

Differential Expression ◽

Differential Methylation ◽

Cpg Methylation ◽

Leukemic Cells ◽

Ctcf Binding ◽

Myeloid Differentiation ◽

Whole Genome

AbstractBackgroundMyeloid differentiation gives rise to a plethora of immune cells in the human body. This differentiation leaves strong signatures in the epigenome through each differentiated state of genetically identical cells. The leukemic HL-60/S4 promyelocytic cell can be easily differentiated from its undifferentiated promyelocyte state into neutrophil-and macrophage-like cell states, making it an excellent system for studying myeloid differentiation. In this study, we present the underlying genome and epigenome architecture of HL-60/S4 through its undifferentiated and differentiated cell states.ResultsWe performed whole genome bisulphite sequencing of HL-60/S4 cells and their differentiated counterparts. With the support of karyotyping, we show that HL-60/S4 maintains a stable genome throughout differentiation. Analysis of differential CpG methylation reveals that most methylation changes occur in the macrophage-like state. Differential methylation of promoters was associated with immune related terms. Key immune genes, CEBPA, GFI1, MAFB and GATA1 showed differential expression and methylation. However, we observed strongest enrichment of methylation changes in enhancers and CTCF binding sites, implying that methylation plays a major role in large scale transcriptional reprogramming and chromatin reorganisation during differentiation. Correlation of differential expression and distal methylation with support from chromatin capture experiments allowed us to identify putative proximal and long-range enhancers for a number of immune cell differentiation genes, including CEBPA and CCNF. Integrating expression data, we present a model of HL-60/S4 differentiation in relation to the wider scope of myeloid differentiation.ConclusionsFor the first time, we elucidate the genome and CpG methylation landscape of HL-60/S4 during differentiation. We identify all differentially methylated regions and positions. We link these to immune function and to important factors in myeloid differentiation. We demonstrate that methylation plays a more significant role in modulating transcription via enhancer reprogramming, rather than by promoter regulation. We identify novel regulatory regions of key components in myeloid differentiation that are regulated by differential methylation. This study contributes another layer of “omics” characterisation of the HL-60/S4 cell line, making it an excellent model system for studying rapid in vitro cell differentiation.Summary statementEpigenomics plays a major role in cell identity and differentiation. We present the DNA methylation landscape of leukemic cells during in-vitro differentiation, to add another ‘omics layer to better understand the mechanisms behind differentiation.

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Regulation of erythropoietin production in a human hepatoblastoma cell line

Blood ◽

10.1182/blood.v70.6.1904.bloodjournal7061904 ◽

1987 ◽

Vol 70 (6) ◽

pp. 1904-1909

Author(s):

OJ Nielsen ◽

SJ Schuster ◽

R Kaufman ◽

AJ Erslev ◽

J Caro

Keyword(s):

Cell Line ◽

Northern Blot ◽

Proliferative Activity ◽

Northern Blot Analysis ◽

Cobalt Chloride ◽

Biologically Active ◽

Erythropoietin Production ◽

Rna Probe ◽

Hepatoblastoma Cell Line

Production of immuno and biologically active erythropoietin was documented to occur in the human hepatoblastoma cell line HepG-2. The expression of the erythropoietin gene was further verified by Northern blot analysis using a single stranded RNA probe. In vitro studies showed that erythropoietin production by these cells was not stimulated by hypoxia or cobalt chloride, but was related to the proliferative activity of the cells in culture. In addition it was found that the secretion of erythropoietin was almost completely abrogated by tunicamycin, an inhibitor of N-linked glycosylation. This effect of tunicamycin was also observed in a permanently transfected cell line that secretes erythropoietin in large quantities.

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