Determination of chromosomal area of double minute chromosomes and a homogeneously staining region in HL-60 human leukemia cells by the use of a color image analyzer


The surface area of chromosomes was measured in two sublines of HL-60 human leukemia cells using a color image analyzer. One subline, HL-60 (dmin), had double minute chromosomes (dmin) and the other line, HL-60 (HSR), showed an abnormal chromosome #8 (8q+) which contained a homogeneously staining region (HSR) within the long arm at band q24. Dmin were not seen in the latter cells. The area of individual chromosomes analyzed from metaphase plates of Giemsa-stained photographic prints proved to be reasonably accurate and reproducible. Moreover, the relative area of each chromosome correlated well both with the relative length of corresponding chromosomes and the relative DNA content. The results indicate that the mean area of a single dmin represents 0.16% of the total genomic area of HL-60 (dmin) cells. The average number of dmin per cell was 8.14, and the total area of all dmin in a representative cell was 1.32% of the entire genome. The 8q+ chromosome had 1.46 times the area of the normal homologous chromosome #8, and the HSR itself had 1.30% of the total chromosomal area of HL-60 (HSR) cells. Thus, perhaps coincidentally, the relative amount of DNA in the HSR in an HL-60 (HSR) cell was similar to the total found in multiple dmin in HL-60 (dmin) cells.


  1. Bahr, G., Gilbert, F., Balaban, G., and Engler, W. 1983. Homogeneously staining regions and double minutes in a human cell line: chromatin organization and DNA content.J. Natl. Cancer Inst. 71: 657–661.

  2. Balaban-Malenbaum, G. and Gilbert, F. 1980. Relationship between homogeneously staining region and double minute chromosomes in human neuroblastoma cell lines. InAdvances in Neuroblastoma Research (A.E. Evans, ed.), Raven Press, New York, pp. 97–107.

  3. Barker, P.E., and Hsu, T.C. 1979. Double minutes in human carcinoma cell lines, with special reference to breast tumors.J. Natl. Cancer Inst. 62: 257–262.

  4. Biedler, J.L. and Spengler, B.A. 1976. A novel chromosome abnormality in human neuroblastoma and antifolate-resistant Chinese hamster cell lines in culture.J. Natl. Cancer Inst. 57: 683–695.

  5. Biedler, J.L., Melera, P.W., and Spengler, B.A. 1980. Specifically altered metaphase chromosomes in antifolate-resistant Chinese hamster cells that overproduce dihydrofolate reductase.Cancer Genet. Cytogenet. 2: 47–60.

  6. Biedler, J.L., Meyers, M.B., and Spengler, B.A. 1983. Homogeneously staining regions and double minute chromosomes, prevalent cytogenetic abnormalities of human neuroblastoma cells.Adv. Cell Neurobiol. 4: 267–307.

  7. Bosman, F.T., van den Ploeg, M., van Duijn, P., and Schaberg, A. 1977. Photometric determination of the DNA distribution in the 24 human chromosomes.Exp. Cell Res. 105: 301–311.

  8. Collins, S.J., Gallo, R.C., and Gallagher, R.E. 1977. Continuous growth and differentiation of human myeloid leukaemic cells in suspension culture.Nature 270: 347–349.

  9. Cowell, J.K. 1982. Double minutes and homogeneously staining regions: Gene amplification in mammalian cells.Ann. Rev. Genet. 16: 21–59.

  10. Cox, D., Yuncken, C., and Springgs, A.I. 1965. Minute chromatin bodies in malignant tumours of childhood.Lancet ii: 55–57.

  11. Fujita, S. 1983. The microcomputer-based color image analyzer and its application to histochemistry.J. Histochem. Cytochem. 31: 238–240.

  12. Gallagher, R.E., Ferrari, A.C., Zulich, A.W., Yen, R.-W.C., and Testa, J.R. 1984. Cytotoxic and cytodifferentiative components of 6-thioguanine resistance in HL-60 cells containing acquired double minute chromosomes.Cancer Res. 44: 2642–2653.

  13. Gallagher, R., Collins, S., Trujillo, J., McCredie, K., Ahearn, M., Tsai, S., Metzgar, R., Aulakh, G., Ting, R., Ruscetti, F., and Gallo, R. 1979. Characterization of the continuous, differentiating myeloid cell line (HL-60) from a patient with acute promyelocytic leukemia.Blood 54: 713–733.

  14. Hamlin, L.J., Milbrandt, J.D., Heintz, N.H., and Azizkhan, J.C. 1984. DNA sequence amplification in mammalian cells.Int. Rev. Cytol. 90: 31–82.

  15. ISCN (1985). 1985.An International System for Human Cytogenetic Nomenclature. Harden D.G. and Klinger, H.P. (eds.); published in collaboration with Cytogenet. Cell Genet. Karger, Basel.

  16. Misawa, S., Staal, S.P., and Testa, J.R. An amplified protooncogene, c-myc, is associated with an abnormally staining region on chromosome 8 or double minute chromosomes in HL-60 human leukemia cells. submitted.

  17. Neel, B.G., Jhanwar, S.C., Chaganti, R.S.K., and Hayward, W.S. 1982. Two human c-onc genes are located on the long arm of chromosome 8.Proc. Natl. Acad. Sci. USA 79: 7842–7846.

  18. Nowell, P., Finan, J., Dalla-Favera, R., Gallo, R.C., ar-Rushdi, A., Romanczuk, H., Selden, J.R., Emanuel, B.S., Rovera, G., and Croce, C.M. 1983. Association of amplified oncogene c-myc with an abnormally banded chromosome 8 in a human leukaemia cell line.Nature 306: 494–497.

  19. Rees, H. and Jones, R.N. 1972. The origin of the wide species variation in nuclear DNA content.Int. Rev. Cytol. 32: 53–92.

  20. Spriggs, A.I., Boddington, M.M., and Clarke, C.M. 1962. Chromosomes of human cancer cells.Br. Med. J. 2: 1431–1435.

  21. Takamatsu, T., Kitamura, T., and Fujita, S. 1986. Quantitative fluorescence image analysis.Acta Histochem. Cytochem. 19: 61–71.

  22. Taub, R., Kirsch, I., Morton, C., Lenoir, G., Swan, D., Tronik, S., Aaronson, S., and Leder, P. 1982. Translocation of the c-myc gene into the immunoglobulin heavy chain locus in human Burkitt lymphoma and murine plasmacytoma cells.Proc. Natl. Acad. Sci. USA 79; 7837–7841.

  23. Wolman, S.R., Lanfrancone, L., Dalla-Favera, R., Ripley, S., and Henderson, A.S. 1985. Oncogene mobility in human leukemia line HL-60.Cancer Genet. Cytogenet. 17: 133–141.

Download references

Author information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Misawa, S., Abe, T., Takamatsu, T. et al. Determination of chromosomal area of double minute chromosomes and a homogeneously staining region in HL-60 human leukemia cells by the use of a color image analyzer. Jap J Human Genet 31, 297–307 (1986).

Download citation


  • Abnormal Chromosome
  • Homologous Chromosome
  • Entire Genome
  • Relative Length
  • Metaphase Plate