Karyotype instability and altered differentiation of rat sarcoma cells after retroviral infection

Steffen, Martín; Scherdin, Ulrich; Hölzel, F.; Vértes, Isabella; Boecker, Werner; Dietel, Manfred

doi:10.1002/gcc.2870040107

Cited by 8 publications

(6 citation statements)

References 49 publications

(43 reference statements)

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“…Our results suggest that inappropriate expression of these meiotic activities in somatic cells interferes with the maintenance of DNA content and add to the in vitro observation that Mos interferes with the ability of PtK1 chromosomes to congress properly (14). These activities would be expected to account for the genetic instability reported for MuSV-infected cells (40 …”

Section: Discussionsupporting

confidence: 56%

Mos overexpression in Swiss 3T3 cells induces meiotic-like alterations of the mitotic spindle.

Fukasawa

Woude

1995

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

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High levels of mos protooncogene product are expressed during oocyte meiotic maturation and Mos has been implicated in formation of the spindle and spindle pole. Here, we show that in Swiss 3T3 cells with 4N DNA content, high levels of Mos lead to the production of binucleated cells. The Swiss 3T3 cells in mitosis, before binucleation occurs, are anastral and the spindle poles are juxtaposed to the cell membrane. These phenotypes may be related to the meiotic process of attachment of the spindle pole to the oocyte membrane during polar body formation. The production of binucleated somatic cells could result from attachment of the altered mitotic spindle pole to the cell membrane that interferes with cytokinesis but not karyokinesis. This can explain at least one form of genetic instability that leads to altered DNA content in tumor cells.In Xenopus, stimulation by progesterone releases stage VI oocytes from G2/M arrest and reinitiates meiotic maturation. Mos expression begins shortly after hormonal stimulation and is required throughout meiosis for the activation, cycling, and maintenance of maturation-promoting factor (1-7). Mos has been shown to be an active component of cytostatic factor (CSF) (8). CSF is believed to arrest oocytes at metaphase II of meiosis by contributing to the stabilization of the meiotic spindle (9) and the germ-line loss of Mos appears to specifically affect female fertility through the failure of maturing oocytes to arrest at metaphase 11 (10,11). Mos has also been implicated in the reorganization of microtubules that leads to formation of the spindle and the spindle pole (12,13) and recently has been shown to associate with kinetochores when microinjected into PtK1 epithelial cells (14). In addition to maturation-promoting factor, Mos activates the mitogenactivated protein kinase (MAPK) cascade (15,16) through the specific activation of MAPK kinase (15,17). MAPK has also been implicated in meiotic microtubule reorganization (18) and, in mouse oocytes, is localized in microtubule organizing centers (19,20). Consistent with the parallels between Mos and MAPK activities and functions, MAPK has been shown to induce CSF activity (21)(22)(23) The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.oocytes during the process of polar body formation. Binucleated cells were previously observed in mouse rhabdomyosarcomas induced by Mo-MuSV (25). Our results provide an explanation for the formation of such binucleated cells that may be related to one form of genetic instability in tumor cells. MATERIALS AND METHODSCells. Swiss 3T3 cells were maintained in complete medium [DMEM/10% fetal bovine serum (FBS) with penicillin (100 units/ml) and streptomycin (100 ,ug/ml)], and grown in an atmosphere containing 10% C02/90% air. Cultures were maintained at a subconfluent density by passing 7.5 x 105 cells per 100-mm dish twice weekly.Virus and V...

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Section: Discussionsupporting

confidence: 56%

Mos overexpression in Swiss 3T3 cells induces meiotic-like alterations of the mitotic spindle.

Fukasawa

Woude

1995

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

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“…4). Because dexamethasone causes partial reversion of some fibrosarcoma cell lines (18,19), we asked whether dexamethasone or indomethacin ( is not activated, suggesting that only a subset of the total cellular response to src is expressed in these LA90 cells.…”

Section: Resultsmentioning

confidence: 99%

Persistent induction of cyclooxygenase in p60v-src-transformed 3T3 fibroblasts.

Han

Sadowski²,

Young³

et al. 1990

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

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A BALB/c 3T3 cell line infected with the temperature-sensitive Rous sarcoma virus strain LA90 has been used to investigate early, p60v"-dependent changes in gene expression (protein synthesis). Giant two-dimensional electrophoresis, which can resolve >3000 polypeptides from[35S]methionine-labeled cell lysates, was used to detect the induction of a p72-74 (72-74 kDa) doublet (pI 7.5) after activation of p60'-at 350C. Antiserum against cyclooxygenase (prostaglandin synthase or prostaglandin endoperoxide synthase) specifically immunoprecipitated the p72-74 doublet. The p72-74 doublet was also induced by platelet-derived growth factor and by phorbol 12-myristate 13-acetate and was elevated in an NIH 3T3 cell line transformed by wild-type src.Activation of p60"-caused a persistant increase in p72-74, whereas the effect of the growth factor was transient. These dissimilar kinetics of induction were paralleled by changes in cyclooxygenase activity. Down-regulation of protein kinase C inhibited subsequent induction of cyclooxygenase by phorbol myristate acetate but did not block induction by p60Nc. The glucocorticoid agonist dexamethasone inhibited induction of cyclooxygenase by p60"-. Although induction of this enzyme may not be directly involved in transformation, the data support the view that oncogenic transformation may result, not from expression of transformation-specific genes, but from persistent changes in the expression of genes normally induced only transiently during passage from the Go stage of the cell cycle.An important goal of studies of oncogene function is to identify gene products, the expression of which may be controlled by the oncogene of interest. Such identification might ultimately determine whether oncogenic transformation (t) is a consequence of the expression of transformationspecific genes, which are not normally expressed in growing cells, or rather (ii) is a result of the continuing expression at inappropriate times of genes normally induced only transiently during mitogenesis. One successful approach to this goal has been through differential and subtractive screening of cDNA libraries prepared from normal and oncogenetransformed cells (1), preferably using temperature-sensitive oncogene mutants; the latter enables the search for immediate-early genes that are induced rapidly rather than as the indirect end product of a cascade of events initiated by the oncogene. This approach has uncovered several interesting genes, the expression of which is altered in primary chicken embryo fibroblasts by the src oncogene (1-4). As an alternative approach to search for src-induced genes, we have exploited the high resolving power of giant two-dimensional electrophoresis (5) to look for rapid changes in protein abundance subsequent to the activation of p60v-src in a BALB/c 3T3 cell line infected with the Rous sarcoma virus temperature-sensitive mutant LA90 of src. The tyrosinespecific protein kinase activity ofthe LA90 p60vsrc is inactive at 40°C but rapidly increases (tl2 = 5 min) when the ce...

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“…The choice of these cell lines was based on two factors: first, the reliability of both cell lines as models has been demonstrated in investigations of the effects of glucocorticoid hormones on cell morphology and proliferation and the stability of cultured rat cells after infection with Moloney murine sarcoma virus [5], [6], [7]; second, these cell lines are commercially available to the entire scientific community, and when they are properly characterized, they may constitute reliable cell models for breast cancer research.…”

Section: Introductionmentioning

confidence: 99%

Defining the Sister Rat Mammary Tumor Cell Lines HH-16 cl.2/1 and HH-16.cl.4 as an In Vitro Cell Model for Erbb2

2012

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Cancer cell lines have been shown to be reliable tools in genetic studies of breast cancer, and the characterization of these lines indicates that they are good models for studying the biological mechanisms underlying this disease. Here, we describe the molecular cytogenetic/genetic characterization of two sister rat mammary tumor cell lines, HH-16 cl.2/1 and HH-16.cl.4, for the first time. Molecular cytogenetic analysis using rat and mouse chromosome paint probes and BAC/PAC clones allowed the characterization of clonal chromosome rearrangements; moreover, this strategy assisted in revealing detected breakpoint regions and complex chromosome rearrangements. This comprehensive cytogenetic analysis revealed an increase in the number of copies of the Mycn and Erbb2 genes in the investigated cell lines. To analyze its possible correlation with expression changes, relative RNA expression was assessed by real-time reverse transcription quantitative PCR and RNA FISH. Erbb2 was found to be overexpressed in HH-16.cl.4, but not in the sister cell line HH-16 cl.2/1, even though these lines share the same initial genetic environment. Moreover, the relative expression of Erbb2 decreased after global genome demethylation in the HH-16.cl.4 cell line. As these cell lines are commercially available and have been used in previous studies, the present detailed characterization improves their value as an in vitro cell model. We believe that the development of appropriate in vitro cell models for breast cancer is of crucial importance for revealing the genetic and cellular pathways underlying this neoplasy and for employing them as experimental tools to assist in the generation of new biotherapies.

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Karyotype instability and altered differentiation of rat sarcoma cells after retroviral infection

Cited by 8 publications

References 49 publications

Mos overexpression in Swiss 3T3 cells induces meiotic-like alterations of the mitotic spindle.

Mos overexpression in Swiss 3T3 cells induces meiotic-like alterations of the mitotic spindle.

Persistent induction of cyclooxygenase in p60v-src-transformed 3T3 fibroblasts.

Defining the Sister Rat Mammary Tumor Cell Lines HH-16 cl.2/1 and HH-16.cl.4 as an In Vitro Cell Model for Erbb2

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