Abstract
A human tumor cell line designated RMS‐GR was established from an embryonal rhabdomyosarcoma. The monolayer cells were polygonal, round or spindle‐shaped. The RMS‐GR cell line became stable with a doubling time of 42 h. Tumorigenicity of the cells was confirmed by hetero‐transplantion into nude mice. Electron microscopic images showed typical cytoplasmic inclusion of aggregated intermediate filaments and myofibril‐like thin filaments. The expression of desmin, vimentin, actin and human myoglobin was recognized by cytofluorometric analyses, and a large fraction of CK‐MM and small fractions of CK‐BB and MCK‐1 isoenzymes were found. Chromosomal analysis showed that the modal chromosome number was consistently near triploid with structural abnormalities mostly involving chromosomes 1, 3 and 8, and additional unidentified markers. No alteration of chromosome 2 was observed. The RMS‐GR cell line may provide a system to identify genes which are involved in the pathogenic mechanism of rhabdomyosarcomas, and to investigate the modulation of myogenic differentiation.
Keywords: Key words, Rhabdomyosarcoma, Intermediate filaments, Actin, Myoglobin, Creatin kinase
Full Text
The Full Text of this article is available as a PDF (995.8 KB).
REFERENCES
- 1. ) Enzinger , F. M. and Weiss , S. W.In“Soft Tissue Tumors,” pp. 539 – 577 ( 1995. ). C.V. Mosby; , St. Louis , MO . [Google Scholar]
- 2. ) Tsokos , M. , Webber , B. L. and Parham , D. M.Rhabdomyosarcoma: a new classification scheme related to prognosis . Arch. Pathol. Lab. , 116 , 847 – 855 ( 1992. ). [PubMed] [Google Scholar]
- 3. ) Newton , W. A. , Gehan , E. A. , Webber , B. L. , Marsden , H. B. , Van Unnik , A. J. , Hamoudi , A. B. , Tsokos , M. G. , Shimada , H. , Harms , D. and Schmidt , D.Classification of rhabdomyosarcoma and related sarcomas . Cancer , 76 , 1073 – 1085 ( 1995. ). [DOI] [PubMed] [Google Scholar]
- 4. ) Tsokos , M.The diagnosis and classification of childhood rhabdomyosarcoma . Semin. Diagn. Pathol. , 11 , 26 – 38 ( 1994. ). [PubMed] [Google Scholar]
- 5. ) Farrell , F. X. , Sax , C. M. , Garzon , R. J. and Zehner , Z. E.Regulation of vimentin expression during myogenesis . In“Cellular and Molecular Biology of Muscle Development,” ed. Kedes L. H. and Stockdale F. E. , pp. 777 – 788 ( 1989. ). Alan R. Liss; , New York . [Google Scholar]
- 6. ) Skalli , O. , Gabbiani , G. , Babai , F. , Seemayer , T. A. , Pizzolato , G. and Schurch , W.Intermediate filament proteins and actin isoforms as markers for soft tissue tumor differentiation and origin . Am. J. Pathol. , 130 , 515 – 531 ( 1988. ). [PMC free article] [PubMed] [Google Scholar]
- 7. ) Schmidt , R. A. , Cone , R. , Haas , J. E. and Grown , A. M.Diagnosis of rhabdomyosarcomas with HHF35, a monoclonal antibody directed against muscle actins . Am. J. Pathol. , 131 , 19 – 28 ( 1988. ). [PMC free article] [PubMed] [Google Scholar]
- 8. ) Kahn , H. J. , Yeger , H. , Kassin , O. , Jorgensen , A. O. , MacLennan , D. , Baumal , R. , Smith , C. R. and Phillips , M. J.Immunohistochemical and electron microscopic assessment of childhood rhabdomyosarcoma . Cancer , 51 , 1897 – 1903 ( 1983. ). [DOI] [PubMed] [Google Scholar]
- 9. ) Apple , F. S. and Billadello , J. S.Expression of creatine kinase M and B mRNAs in treadmill trained rat skeletal muscle . Life Sci. , 55 , 558 – 592 ( 1994. ). [DOI] [PubMed] [Google Scholar]
- 10. ) Moss , D. W. , Whitaker , K. B. and Munro , A. J.Creatine kinase MB isoenzyme in rhabdomyosarcoma . Clin. Chem. , 35 , 896 – 897 ( 1989. ). [PubMed] [Google Scholar]
- 11. ) Fernández , J. E. , Rodríguez , F. , Recio , M. A. , Vélez , C. , Aránega , A. E. , Alvarez , L. and Aránega , A.Detection of creatine kinase isoenzymes as tumoral markers of rhabdomyosarcoma . Enzyme , 46 , 245 – 248 ( 1992. ). [DOI] [PubMed] [Google Scholar]
- 12. ) Sreekantaiah , C. , Ladanyi , M. , Rodriguez , E. and Chaganti , R. S. K.Chromosomal aberrations in soft tissue tumours. Relevance to diagnosis, classification and molecular mechanisms . Am. J. Pathol. , 144 , 1121 – 1134 ( 1994. ). [PMC free article] [PubMed] [Google Scholar]
- 13. ) Parham , D. M. , Shapiro , D. N. , Downing , J. R. , Webber , B. L. and Douglass , E. C.Solid alveolar rhabdomyosarcomas with the t(2;13) . Am. J. Surg. Pathol. , 18 , 474 – 478 ( 1994. ). [DOI] [PubMed] [Google Scholar]
- 14. ) Trent , J. , Casper , J. , Meltzer , P. , Thomson , F. and Fogh , J.Nonrandom chromosome alterations in rhabdomyosarcoma . Cancer Genet. Cytogenet. , 16 , 189 – 192 ( 1985. ). [DOI] [PubMed] [Google Scholar]
- 15. ) Fernández , J. E. , Concha , A. , Aránega , A. , Ruiz‐Cabello , F. , Cabrera , T. and Garrido , F.HLA Class I and II expression in rhabdomyosarcoma . Immunobiology , 182 , 440 – 448 ( 1991. ). [DOI] [PubMed] [Google Scholar]
- 16. ) Seabright , M.A rapid banding technique for human chromosomes . Lancet , ii , 971 ( 1971. ). [DOI] [PubMed] [Google Scholar]
- 17. ) Garvin , A. J. , Stanley , W. S. , Bennet , D. D. , Sullivan , J. L. and Sens , D. A.The in vitro growth, heterotransplantation, and differentiation of a human rhabdomyosarcoma cell line . Am. J. Pathol. , 125 , 208 – 217 ( 1986. ). [PMC free article] [PubMed] [Google Scholar]
- 18. ) Hayashi , Y. , Sugimoto , T. , Horii , Y. , Hosoi , H. , Inazawa , J. , Kemshead , J. T. , Inaba , T. , Hanada , R. , Yamamoto , K. and Gown , A. M.Characterization of an embryonal rhabdomyosarcoma cell line showing amplification and overexpression of the N‐myc oncogene . Int. J. Cancer , 45 , 705 – 711 ( 1990. ). [DOI] [PubMed] [Google Scholar]
- 19. ) Kubo , K. , Naoe , T. , Utsumi , K. R. , Ishiguro , Y. , Ueda , K. , Shiku , H. and Yamada , K.Cytogenetic and cellular characteristics of a human embryonal rhabdomyosarcoma cell line, RMS‐YM . Br. J. Cancer , 63 , 879 – 884 ( 1991. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 20. ) Karnes , P. S. , Tran , T. N. , Cui , M. Y. , Bogenmann , E. , Shimada , F. , Shimada , H. and Ying , K. L.Establishment of a rhabdoid tumor cell line with a specific chromosomal abnormality, 46, XY, f/11;22/(p15,5;q11.23) . Cancer Genet. Cytogenet. , 56 , 31 – 38 ( 1991. ). [DOI] [PubMed] [Google Scholar]
- 21. ) Kanako , S. , Kanako , M. and Ohkawa , H.Establishment and characterization of a childhood embryonal rhabdomyosarcoma cell line with nonrandom chromosome alterations . Human Cell , 8 , 35 – 41 ( 1995. ). [PubMed] [Google Scholar]
- 22. ) Aguanno , S. , Bouche , M. , Sergio , A. and Molinaro , M.12‐O‐Tetradecanoylphorbol‐13‐acetate‐induced differentiation of a human rhabdomyosarcoma cell line . Cancer Res. , 50 , 3377 – 3382 ( 1990. ). [PubMed] [Google Scholar]
- 23. ) Prados , J. , Melguizo , C. , Fernández , J. E. , Aránega , A. E. , Alvarez , L. and Aránega , A.Actin, tropomyosin and α‐actinin as markers of differentation in human rhabdomyosarcoma cell lines induced with dimethyl sulfoxide . Cell. Mol. Biol. , 39 , 525 – 536 ( 1993. ). [PubMed] [Google Scholar]
- 24. ) Ramp , U. , Gerharz , C. D. , Engers , R. , Marx , N. and Gabbert , H.E.Differentiation induction in the human rhabdomyosarcoma cell line Te‐671‐A. Morphological, biochemical and molecular analysis . Anticancer Res. , 15 , 181 – 188 ( 1995. ). [PubMed] [Google Scholar]
- 25. ) Shapiro , D. N. , Houghton , P. J. , Hazelton , B. J. , Germain , G. S. , Murti , K. G. , Rahman , A. and Houghton , J. A.Morphological and molecular characterization of spontaneous myogenic differentiation in a human rhabdomyosarcoma cell line . Cancer Res. , 50 , 6002 – 6009 ( 1990. ). [PubMed] [Google Scholar]
- 26. ) Petkovi , C. I. , Dominis , M. , Nakic , M. and Cepulic , M.Embryonal rhabdomyosarcoma with 92, XXYY+dmin and evidence of spontaneous cell fusion . Cancer Genet. Cytogenet. , 76 , 125 – 128 ( 1994. ). [DOI] [PubMed] [Google Scholar]
- 27. ) Melguizo , C. , Prados , J. , Velez , C. , Aranega , A. E. , Alvarez , L. and Aranega , A.Influence of dimethyl sulfoxide on intermediate filament proteins in human rhabdomyosarcoma cell lines: modulation at subcellular level . Histochem. J. , 26 , 519 – 525 ( 1994. ). [DOI] [PubMed] [Google Scholar]
- 28. ) Perriard , J. C. , Hossle , J. P. , Schäfer , B. W. , Soldati , T. , Wegmann , G. and Wirz , T.Multiple tasks of creatine kinase isoproteins . In“Cellular and Molecular Biology,” ed. Kedes L. H. and Stockdale F. E. , pp. 545 – 554 ( 1989. ). Alan R. Liss; , New York . [Google Scholar]
- 29. ) Tsokos , M. , Howar , R. and Costa , J.Immunohistochemical study of alveolar and embryonal rhabdomyosarcoma . Lab. Invest. , 48 , 148 – 155 ( 1983. ). [PubMed] [Google Scholar]
- 30. ) de Jong , A. S. H. , Van Kessel‐Van Vark , M. , Albus‐Lutter , C. E. and Voute , P.A.Creatine kinase subunits M and B as markers in the diagnosis of poorly differentiated rhabdomyosarcomas in children . Hum. Pathol. , 16 , 924 – 928 ( 1985. ). [DOI] [PubMed] [Google Scholar]
- 31. ) Fernández , J. E. , Prados , J. , Melguizo , C. , Rodriguez , F. , Alvarez , L. and Aranega , A.Creatine kinase isoenzymes as markers of differentiation in rhabdomyosarcoma cells in culture . Pathol. Res. Pract. , 190 , 1169 – 1173 ( 1994. ). [DOI] [PubMed] [Google Scholar]
- 32. ) Barr , F. G. , Holick , J. , Nycum , L. , Biegel , J. A. and Emanuel , B.S.Localization of the t(2;13) breakpoint of alveolar rhabdomyosarcoma on a physical map of chromosome 2 . Genomics , 13 , 1150 – 1156 ( 1992. ). [DOI] [PubMed] [Google Scholar]
- 33. ) Macina , R. A. , Barr , F. G. , Galili , N. and Riethman , H. C.Genomic organization of the human PAX3 gene: DNA sequence analysis of the region disrupted in alveolar rhabdomyosarcoma . Genomics , 26 , 1 – 8 ( 1995. ). [DOI] [PubMed] [Google Scholar]
- 34. ) Ogose , A. , Motoyama , T. , Hotta , T. and Watanche , H.In vitro differentiation and proliferation in a newly established human rhabdomyosarcoma cell line . Virchows Arch. , 426 , 385 – 391 ( 1995. ). [DOI] [PubMed] [Google Scholar]
- 35. ) Wang‐Wuu , S. , Soukup , S. , Ballard , E. , Gotwals , B. and Lampkin , B.Chromosomal analysis of sixteen human rhabdomyosarcomas . Cancer Res. , 48 , 983 – 987 ( 1988. ). [PubMed] [Google Scholar]
- 36. ) Douglass , E. C. , Green , A. A. , Hayes , F. A. , Etaubanas , E. , Horowitz , M. and Williams , J. A.Chromosome 1 abnormalities: a common feature of pediatric solid tumors . J. Natl. Cancer Inst. , 75 , 51 – 55 ( 1985. ). [PubMed] [Google Scholar]
- 37. ) Magnani , I. , Faustirella , F. , Nanni , P. , Nicoletti , G. and Larizza , L.Karyotypic characterization of a new embryonal rhabdomyosarcoma cell line . Cancer Genet. Cytogenet , 54 , 83 – 89 ( 1991. ). [DOI] [PubMed] [Google Scholar]