Abstract
The process of metastasis was analyzed in 505‐05‐01 cells, an established line of a methylcholanthrene‐induced mouse sarcoma, by tagging the cells with the pSV2neo plasmid. A dominant clone was identified which, upon transplantation together with other clones, overgrew tumors in the kidney capsule. However, when this clone was transplanted in a mouse collaterally with recessive clones, both clones grew at the same rate and metastasized to the lung at an equal frequency. This suggests that the process of metastasis in this particular sarcoma line is stochastic, the dominant population having a better chance to colonize to the lung. The dominant neomycin‐resistant clone was transfected with another marker plasmid, pY3, which confers resistance to hygromycin. Results of mixed inoculation of 9 independently isolated clones revealed the hierarchy of dominance among clones. This indicated the existence of heterogeneity within the parental clone. Upon mixed inoculation with hygromycin‐resistant clones, the parental clone overgrew in the tumors. This indicated that some clone had changed its phenotype to become less aggressive. Thus, the direction of phenotypic drift in vitro seems to be random in terms of behavior in vivo.
Keywords: Gene tagging, Stochastic metastasis, Phenotypic instability, Random drift
Full Text
The Full Text of this article is available as a PDF (484.7 KB).
REFERENCES
- 1. ) Rubin , H.The significance of biological heterogeneity . Cancer Metastasis Rev. , 9 , 1 – 20 ( 1990. ). [DOI] [PubMed] [Google Scholar]
- 2. ) Tamura , G. , Kihana , T. , Nomura , K. , Terada , M. , Sugimura , T. and Hirohashi , S.Detection of frequent p53 gene mutations in primary gastric cancer by cell sorting and polymerase chain reaction single‐strand conformation polymorphism analysis . Cancer Res. , 51 , 3056 – 3058 ( 1991. ). [PubMed] [Google Scholar]
- 3. ) Tlsty , T. , White , A. and Sanchez , J.Suppression of gene amplification in human cell hybrids . Science , 255 , 1425 – 1427 ( 1992. ). [DOI] [PubMed] [Google Scholar]
- 4. ) Hastie , N. D. , Dempster , M. , Dunlop , M. G. , Thompson , A. M. , Green , D. K. and Allshire , R. C.Telomere reduction in human colorectal carcinoma and with aging . Nature , 346 , 866 – 868 ( 1990. ). [DOI] [PubMed] [Google Scholar]
- 5. ) Kuchino , Y. , Mori , F. , Kasai , H. , Inoue , H. , Iwai , S. , Miura , K. , Ohtsuka , E. and Nishimura , S.Misreading of DNA templates containing 8‐hydroxydeoxyguanosine at the modified base and at adjacent residues . Nature , 327 , 77 – 79 ( 1987. ). [DOI] [PubMed] [Google Scholar]
- 6. ) Szatrowski , T. and Nathan , C. F.Production of large amounts of hydrogen peroxide by human tumor cells . Cancer Res. , 51 , 794 – 798 ( 1991. ). [PubMed] [Google Scholar]
- 7. ) Malins , D. C. and Haimanot , R.Major alterations in the nucleotide structure of DNA in cancers of the female breast . Cancer Res. , 51 , 5430 – 5432 ( 1991. ). [PubMed] [Google Scholar]
- 8. ) Fidler , I. J. and Hart , I. R.Biological diversity in metastatic neoplasms: origins and implications . Science , 217 , 998 – 1003 ( 1982. ). [DOI] [PubMed] [Google Scholar]
- 9. ) Liotta , L. and Steler‐Stevenson , W. G.Tumor invasion and metastasis: an imbalance of positive and negative regulation . Cancer Res., (Suppl.) , 51 , 5054 – 5059 ( 1991. ). [PubMed] [Google Scholar]
- 10. ) Nowell , P. C.Mechanism of tumor progression . Cancer Res. , 46 , 2203 – 2207 ( 1986. ). [PubMed] [Google Scholar]
- 11. ) Wainscoat , J. S. and Fey , M. F.Assessment of clonality in human tumors: a review . Cancer Res. , 50 , 1355 – 1360 ( 1990. ). [PubMed] [Google Scholar]
- 12. ) Enoki , Y. , Niwa , O. , Yokoro , K. and Toge , T.Analysis of clonal evolution in a tumor consisting of pSV2neotransfected mouse fibrosarcoma clones . Jpn. J. Cancer Res. , 81 , 141 – 147 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 13. ) Kodama , Y. , Seyama , T. and Yokoro , K.Establishment and immunologic characterization of 3‐methylcholanthrene‐induced sarcoma lines metastasizing widely in mice and exhibiting distinct and selective propensities for the mode of metastasis . J. Natl. Cancer Inst. , 69 , 595 – 605 ( 1982. ). [PubMed] [Google Scholar]
- 14. ) Tsukiyama , T. , Niwa , O. and Yokoro , K.Mechanism of suppression of the long terminal repeat of Moloney leukemia virus in mouse embryonal carcinoma cells . Mol. Cell. Biol. , 9 , 4670 – 4676 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 15. ) Blochlenger , K. and Diggelmann , H.Hygromycin B phosphotransferase as a selectable marker for DNA transfer experiments with higher eukaryotic cells . Mol. Cell. Biol. , 4 , 2929 – 2931 ( 1984. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 16. ) Niwa , O. , Enoki , Y. and Yokoro , K.Overexpression and amplification of the c‐myc gene in mouse tumors induced by chemicals and radiations . Jpn. J. Cancer Res. , 80 , 212 – 218 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 17. ) Talmadge , C. , Tanio , Y. , Meeker , A. , Talmadge , J. and Zbar , B.Tumor cells transfected with the neomycin resistance gene (neo) contain unique genetic markers useful for identification of tumor recurrence and metastasis . Invasion Metastasis , 7 , 197 – 207 ( 1987. ). [PubMed] [Google Scholar]
- 18. ) Korczak , B. , Robson , I. , Lamarche , C. , Bernstein , A. , and Kerbel , R.Genetic tagging of tumor cells with retrovirus vectors: clonal analysis of tumor growth and metastasis in vivo . Mol Cell. Biol. , 8 , 3143 – 3149 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 19. ) Talmadge , J. E. and Fidler , I. J.Cancer metastasis is selective or random depending on the parental tumor population . Nature , 297 , 593 – 594 ( 1982. ). [DOI] [PubMed] [Google Scholar]
- 20. ) Kerbel , R. S. , Waghorne , C. , Man , M. S. , Elliott , B. and Breitman , M. L.Alteration of the tumorigenic and metastatic properties of neoplastic cells is associated with the process of calcium phosphate‐mediated DNA transfection . Proc. Natl. Acad. Sci. USA , 84 , 1263 – 1267 ( 1987. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 21. ) Takada , T. , Suzuki , S. , Sugawara , Y. , Kominami , R. , Arakawa , M. , Niwa , O. and Yokoro , K.Somatic mutation during metastasis of a mouse fibrosarcoma line detected by DNA fingerprint analysis . Jpn. J. Cancer Res. , 83 , 165 – 170 ( 1992. ). [DOI] [PMC free article] [PubMed] [Google Scholar]