Synthesis of Mouse Haemoglobin and Globin mRNA in Leukaemic Cell Cultures

Ostertag, Wolfram; Melderis, H.; Steinheider, G.; Kluge, N.; Dube, Shyam K.

doi:10.1038/newbio239231a0

Cited by 208 publications

(88 citation statements)

References 12 publications

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“…The regulatory mechanism that controls the Hb F to Hb A switch is not known, Murine erythroleukemic cells (MELC) have been used as an in vitro model to start erythroid differentiation [2,3]. The hemoglobins synthesized in the clones derived from the DBA/2 mouse are indistinguishable from the host animal [4,5]. The DBA/2 adult has two hemoglobins which have been designated p-major and P-minor hemoglobins 161.…”

Section: Introdu~ionmentioning

confidence: 99%

“…The DBA/2 adult has two hemoglobins which have been designated p-major and P-minor hemoglobins 161. After induction of MELC with d~e~ylsu~oxide (DMSO) [4,, butyric acid (BA) [8-lo], or hemin [9,1 l-131, the different types of hemo~ob~s are synthesized in different quantities. Therefore, the clones derived from the DBA/2 mouse have been postulated as a useful model system to study gene switching [9].…”

Section: Introdu~ionmentioning

confidence: 99%

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Mouse fetal hemoglobin synthesis in murine erythroleukemic cells

Wu¹,

Zucker²

1979

FEBS Letters

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Section: Introdu~ionmentioning

confidence: 99%

Section: Introdu~ionmentioning

confidence: 99%

Mouse fetal hemoglobin synthesis in murine erythroleukemic cells

Wu¹,

Zucker²

1979

FEBS Letters

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“…The present study examines the relationship between the cell cycle and the expression of a new pattern of genetic information in progeny of cells induced to differentiate (1). The murine erythroleukemia cell (MELC) line established by Friend et al (2) shows a low level (<1%) of spontaneous erythroid differentiation in culture; inclusion of 280 mM dimethyl sulfoxide (Me2SO) in the medium causes a high proportion of the cells to express a program of erythroid differentiation, including morphological changes (2), accumulation of globin mRNAs (3)(4)(5)(6), a and f globin synthesis (7), increase in heme synthesis (8), synthesis of erythrocyte-specific proteins (9), loss in capacity for cell division (2,10), and the appearance of erythrocytespecific membrane proteins (11).…”

mentioning

confidence: 99%

Transient inhibition of initiation of S-phase associated with dimethyl sulfoxide induction of murine erythroleukemia cells to erythroid differentiation.

Terada¹,

Fried²,

Nudel³

et al. 1977

Proc. Natl. Acad. Sci. U.S.A.

108

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The murine erythroleukemia cell (MELC) line in suspension culture can be induced to differentiate to erythroid cells by various compounds, including dimethyl sulfoxide (Me2SO). Analysis of the cell cycle, during differentiation induced by Me2SO, using thymidine incorporation, thymidine labeling index, and relative DNA content per cell as measured by flow microfluorometry, demonstrates a transient inhibition of entry of (2), accumulation of globin mRNAs (3-6), a and f globin synthesis (7), increase in heme synthesis (8), synthesis of erythrocyte-specific proteins (9), loss in capacity for cell division (2, 10), and the appearance of erythrocytespecific membrane proteins (11).The rate of DNA synthesis, proportion of cells in S-phase, and the pattern of DNA accumulation have been measured and compared in MELC grown in the presence and absence of Me2SO and other inducing agents, e.g., butyric acid (12) and dimethylacetamide (13). DNA synthesis has been measured using incorporation of isotopically labeled thymidine in pulse-labeling experiments; cells in S-phase were assayed by autoradiography of thymidine pulse-labeled cells; DNA accumulation per cell was determined using propidium iodide staining with flow microfluorometric analysis. By MATERIALS AND METHODS Cells. Strain 745-A, which is infected with Friend virus complex, was kindly provided by Charlotte Friend and has been maintained in culture in our laboratory as described elsewhere (14). Isolation of the Me2SO-sensitive cell line (DS19) and resistant cell line (DR10) was described by Ohta et al. (15). Cell concentration and proportion of hemoglobin-containing (benzidine-reactive) cells were determined as previously described (14).Pulse Labeling with Thymidine. DNA synthesis was determined as incorporation of [3Hjthymidine (20 Ci/mmol, New England NuclearCorp.) into cells (2 X 105 cells/per ml) incubated for 20 min at 370 at a concentration of 20 ,Ci/ml of culture medium. Thymidine pulse labeling in this manner was performed on aliquots removed from culture at the times indicated for each experiment. Following incubation with radioisotope, cells were washed with phosphate-buffered saline, pH 7.4, containing 2 mM thymidine, and precipitated with 10% trichloroacetic acid. Precipitates were collected on Millipore filters and their radioactivity was measured in Aquasol. A separate aliquot of cell suspension was removed from the culture and, washed with the same phosphate-buffered saline, and smears were prepared employing the cytocentrifuge. These slides were processed for radioautography (16

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“…The tumours induced by us differed; from those induced in DBA/2 mice by other investigators in that the latter continued to relicate virus, differentiated along the erthyrocytic line, and were able to incorporate radioactive iron and synthesize haemoglobin Scher, Ostertag et al, 1972). When dimethylsulphoxide (DMSO) was added to these cultures, the number of cells maturing along the erythrocytic line increased greatly, with proerythroblasts maturing to normoblasts.…”

mentioning

confidence: 58%

Mechanism for the incorporation of S-(1,2,3,4-tetrahydro-2-hydroxy-1-naphthyl)-L-cysteine into protein

Morrison¹,

Whybrew²,

Sobhy³

et al. 1977

Br J Cancer

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Summary.-A transplantable reticulum-cell sarcoma induced by Rauscher virus (RV) in (C57BL/6 x DBA/2)F1 (BDF1) mice was grown in tissue culture. Four separate cell lines were established, all of which grew predominantly in suspension. The doubling time of the cells from these cultures ranged from 17 to 32 h. Each culture continued to replicate RV, as indicated by the infectivity in newborn mice of all fluids tested up to the 75th passage. Since the morphological appearance of the cells in vitro was consistent with that of proerythroblasts, all cultures were tested for their ability to differentiate along the erythrocytic line under the influence of dimethylsulphoxide (DMSO). One of the cultures produced small quantities of haemoglobin independently of DMSO. Another was shown to produce haemoglobin, as well as to take up 59Fe and incorporate it into haem, only in the presence of DMSO. The 2 remaining cultures failed to produce haemoglobin, either spontaneously or in the presence of DMSO. Cells from each of the RV-induced cultures, when inoculated back into BDF1 mice, induced typical reticulum-cell sarcomas, without in vivo evidence of erythroid differentiation. In contrast, 2 morphologically identical but non-infectious cell lines derived from Friend virus-induced reticulumcell sarcomas did not show erythroid differentiation in vivo or in vitro, either in the absence or presence of DMSO.

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Synthesis of Mouse Haemoglobin and Globin mRNA in Leukaemic Cell Cultures

Cited by 208 publications

References 12 publications

Mouse fetal hemoglobin synthesis in murine erythroleukemic cells

Mouse fetal hemoglobin synthesis in murine erythroleukemic cells

Transient inhibition of initiation of S-phase associated with dimethyl sulfoxide induction of murine erythroleukemia cells to erythroid differentiation.

Mechanism for the incorporation of S-(1,2,3,4-tetrahydro-2-hydroxy-1-naphthyl)-L-cysteine into protein

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