Morphological and molecular characterization of spontaneous myogenic differentiation in a human rhabdomyosarcoma cell line.

Skeletal muscle differentiation consists of an ordered withdrawal of committed cells from the cell cycle and their fusion to form multinucleated myotubes. To determine if differentiation of malignant myoblasts parallels that of normal skeletal muscle, a cell line (Rh28) was established from an alveolar rhabdomyosarcoma. Rh28 displays a constant population doubling time of 45-55 h until passage 60, when the doubling time progressively increases until proliferation ceases. Loss of proliferative capacity is associated with morphological evidence of differentiation to multinucleated myotubes, fusion, and the expression of numerous muscle-specific genes. In contrast to normal myogenic differentiation, multinucleated cells continue to synthesize DNA and express abundant c-myc transcripts. These observations suggest synchronous replication and possible arrest in the G2-phase of the cell cycle, since there was no evidence of mitotic activity in differentiated cells. Terminal differentiation of early passage Rh28 cells was induced in the presence of 10% dialyzed fetal calf serum but not by medium containing 2% undialyzed serum, suggesting a role for low molecular weight growth factors in this process. Our data indicate that the Rh28 cell line may be of value in elucidating the relationship between oncogenic transformation and differentiation in rhabdomyosarcoma.

[1]  T. Braun,et al.  Differential expression of myogenic determination genes in muscle cells: possible autoactivation by the Myf gene products. , 1989, The EMBO journal.

[2]  P. Lollini,et al.  Myogenic differentiation of human rhabdomyosarcoma cells induced in vitro by antineoplastic drugs. , 1989, Cancer research.

[3]  E. Olson,et al.  A gene with homology to the myc similarity region of MyoD1 is expressed during myogenesis and is sufficient to activate the muscle differentiation program. , 1989, Genes & development.

[4]  E. Taparowsky,et al.  Inhibition of myogenic differentiation by the H-ras oncogene is associated with the down regulation of the MyoD1 gene. , 1989, Oncogene.

[5]  David Baltimore,et al.  A new DNA binding and dimerization motif in immunoglobulin enhancer binding, daughterless, MyoD, and myc proteins , 1989, Cell.

[6]  T. Braun,et al.  A novel human muscle factor related to but distinct from MyoD1 induces myogenic conversion in 10T1/2 fibroblasts. , 1989, The EMBO journal.

[7]  Victor K. Lin,et al.  Myogenin, a factor regulating myogenesis, has a domain homologous to MyoD , 1989, Cell.

[8]  D. Kelvin,et al.  Growth factors, signaling pathways, and the regulation of proliferation and differentiation in BC3H1 muscle cells. II. Two signaling pathways distinguished by pertussis toxin and a potential role for the ras oncogene , 1989, The Journal of cell biology.

[9]  B. Gallie,et al.  A sensitive method for immunocytochemical detection of P-glycoprotein in multidrug-resistant human ovarian carcinoma cell lines. , 1988, Laboratory investigation; a journal of technical methods and pathology.

[10]  K. Latham,et al.  Myogenic lineage determination and differentiation: Evidence for a regulatory gene pathway , 1988, Cell.

[11]  Torti,et al.  Tumor necrosis factor inhibits human myogenesis in vitro , 1988, Molecular and cellular biology.

[12]  R. Moll,et al.  Terminally differentiated postmitotic tumor cells in a rat rhabdomyosarcoma cell line , 1988, Virchows Archiv. B, Cell pathology including molecular pathology.

[13]  H. Weintraub,et al.  Expression of a single transfected cDNA converts fibroblasts to myoblasts , 1987, Cell.

[14]  H. Blau,et al.  Differential patterns of transcript accumulation during human myogenesis , 1987, Molecular and cellular biology.

[15]  G. Spizz,et al.  Inhibition of myogenic differentiation by fibroblast growth factor or type beta transforming growth factor does not require persistent c-myc expression. , 1987, Developmental biology.

[16]  N. Denis,et al.  c-myc oncogene expression inhibits the initiation of myogenic differentiation. , 1987, Experimental cell research.

[17]  P. Houghton,et al.  Characterization of cell lines derived from xenografts of childhood rhabdomyosarcoma. , 1987, Cancer research.

[18]  G. Spizz,et al.  The oncogenic forms of N-ras or H-ras prevent skeletal myoblast differentiation , 1987, Molecular and cellular biology.

[19]  G. Spizz,et al.  Autonomous expression of c-myc in BC3H1 cells partially inhibits but does not prevent myogenic differentiation , 1987, Molecular and cellular biology.

[20]  J. Chirgwin,et al.  Isolation of RNA using guanidinium salts. , 1987, Methods in enzymology.

[21]  P. Houghton,et al.  A specific chromosomal abnormality in rhabdomyosarcoma. , 1987, Cytogenetics and cell genetics.

[22]  P. Lollini,et al.  RMZ: a new cell line from a human alveolar rhabdomyosarcoma. In vitro expression of embryonic myosin. , 1986, British Journal of Cancer.

[23]  H. Blau,et al.  Insulin and insulinlike growth factor receptors and responses in cultured human muscle cells. , 1986, The American journal of physiology.

[24]  J. Garson,et al.  N-myc ONCOGENE AMPLIFICATION IN RHABDOMYOSARCOMA AT RELEASE , 1986, The Lancet.

[25]  S. Schiaffino,et al.  Embryonic myosin heavy chain as a differentiation marker of developing human skeletal muscle and rhabdomyosarcoma. A monoclonal antibody study. , 1986, Experimental cell research.

[26]  T. Kao,et al.  Isolation and characterization of rat and human glyceraldehyde-3-phosphate dehydrogenase cDNAs: genomic complexity and molecular evolution of the gene. , 1985, Nucleic acids research.

[27]  W. Quax,et al.  Intermediate filament cDNAs from BHK-21 cells: demonstration of distinct genes for desmin and vimentin in all vertebrate classes. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[28]  W. B. Butler,et al.  Preparing nuclei from cells in monolayer cultures suitable for counting and for following synchronized cells through the cell cycle. , 1984, Analytical biochemistry.

[29]  I. Verma,et al.  Expression of cellular oncogenes in human malignancies. , 1984, Science.

[30]  W. Claycomb,et al.  Acquisition of multiple nuclei and the activity of DNA polymerase alpha and reinitiation of DNA replication in terminally differentiated adult cardiac muscle cells in culture. , 1983, Developmental biology.

[31]  A. Feinberg,et al.  A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. , 1983, Analytical biochemistry.

[32]  K. Murti,et al.  Interaction of frog virus-3 with the cytoskeleton. I. Altered organization of microtubules, intermediate filaments, and microfilaments , 1983, The Journal of cell biology.

[33]  H. Varmus,et al.  Homogeneously staining chromosomal regions contain amplified copies of an abundantly expressed cellular oncogene (c-myc) in malignant neuroendocrine cells from a human colon carcinoma. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[34]  D. Alcorta,et al.  Cytoskeletal F-actin patterns quantitated with fluorescein isothiocyanate-phalloidin in normal and transformed cells. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[35]  B. Nadal-Ginard,et al.  Commitment, fusion and biochemical differentiation of a myogenic cell line in the absence of DNA synthesis , 1978, Cell.

[36]  R. Devlin,et al.  Coordinate regulation of contractile protein synthesis during myoblast differentiation , 1978, Cell.

[37]  J. Merlie,et al.  Molecular aspects of myogenesis. , 1977, Current topics in developmental biology.

[38]  E. Lazarides,et al.  Actin, alpha-actinin, and tropomyosin interaction in the structural organization of actin filaments in nonmuscle cells , 1976, The Journal of cell biology.