Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1969 Nov 1;43(2):263–274. doi: 10.1083/jcb.43.2.263

CONTROL OF PIGMENT PRODUCTION IN MOUSE MELANOMA CELLS IN VITRO

Evocation and Maintenance

Selma Silagi 1
PMCID: PMC2107859  PMID: 4981070

Abstract

A clonally derived amelanotic melanoma cell line repeatedly has been forced to produce pigment by the inhibitor of DNA synthesis, I-β-D-arabinofuranosylcytosine (ara-C) at sublethal levels. One ara-C-derived melanotic line has been cloned, and has continued to produce pigment for 2 years on normal medium. The inhibitor is most effective when administered to synchronized cells in four pulses on successive days at 1.8 x 10-5 M during the S phase of the cell cycle. Colcemid at a sublethal concentration, and growth on medium solidified with agar also evoked pigment production in this line, but a large number of other inhibitors of biosynthetic processes did not, under the conditions tested. The melanotic lines are active producers of tyrosinase (DOPA oxidase), whereas the amelanotic line produces an inhibitor of tyrosinase activity. Both enzyme and inhibitor are labile at 4° C and -20° C, and decay of the inhibitor in homogenates of amelanotic cells reveals a low level of residual DOPA oxidase activity. The mean population doubling time of a cloned melanotic line is 23 hr, and that of a cloned amelanotic line 16.5 hr. A similar decrease in rate of growth is found in other melanotic lines and is believed to be a significant factor in maintaining this differentiated function. Rapid growth may be related to the production of an inhibitor by the amelanotic cells.

Full Text

The Full Text of this article is available as a PDF (820.1 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Burnett J. B., Seiler H., Brown I. V. Separation and characterization of multiple forms of tyrosinase from mouse melanoma. Cancer Res. 1967 May;27(5):880–889. [PubMed] [Google Scholar]
  2. CHU M. Y., FISCHER G. A. A proposed mechanism of action of 1-beta-D-arabinofuranosyl-cytosine as an inhibitor of the growth of leukemic cells. Biochem Pharmacol. 1962 Jun;11:423–430. doi: 10.1016/0006-2952(62)90225-3. [DOI] [PubMed] [Google Scholar]
  3. Cahn R. D., Cahn M. B. Heritability of cellular differentiation: clonal growth and expression of differentiation in retinal pigment cells in vitro. Proc Natl Acad Sci U S A. 1966 Jan;55(1):106–114. doi: 10.1073/pnas.55.1.106. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cohen S. S. Introduction to the biochemistry of D-arabinosyl nucleosides. Prog Nucleic Acid Res Mol Biol. 1966;5:1–88. doi: 10.1016/s0079-6603(08)60231-7. [DOI] [PubMed] [Google Scholar]
  5. Coon H. G., Cahn R. D. Differentiation in vitro: effects of Sephadex fractions of chick embryo extract. Science. 1966 Sep 2;153(3740):1116–1119. doi: 10.1126/science.153.3740.1116. [DOI] [PubMed] [Google Scholar]
  6. EAGLE H. Amino acid metabolism in mammalian cell cultures. Science. 1959 Aug 21;130(3373):432–437. doi: 10.1126/science.130.3373.432. [DOI] [PubMed] [Google Scholar]
  7. EPHRUSSI B., TEMIN H. M. Infection of chick iris epithelium with the Rous sarcoma virus in vitro. Virology. 1960 Jul;11:547–552. doi: 10.1016/0042-6822(60)90099-4. [DOI] [PubMed] [Google Scholar]
  8. FOSTER M., TREMBLY G. S., STAMAS T. A. A turbidimetric assay for tyrosinase activity in frozen sections of mouse skin. Proc Soc Exp Biol Med. 1954 Jun;86(2):313–315. doi: 10.3181/00379727-86-21083. [DOI] [PubMed] [Google Scholar]
  9. GREEN H., GOLDBERG B. KINETICS OF COLLAGEN SYNTHESIS BY ESTABLISHED MAMMALIAN CELL LINES. Nature. 1963 Dec 14;200:1097–1098. doi: 10.1038/2001097a0. [DOI] [PubMed] [Google Scholar]
  10. HIRSCH H. M., ZELICKSON A. S. AN ENZYMATIC AND ELECTRON MICROSCOPIC CHARACTERIZATION OF A VARIANT OF THE CLOUDMAN S-91 MELANOMA. Cancer Res. 1964 Aug;24:1137–1153. [PubMed] [Google Scholar]
  11. HU F., LESNEY P. F. THE ISOLATION AND CYTOLOGY OF TWO PIGMENT CELL STRAINS FROM B-16 MOUSE MELANOMAS. Cancer Res. 1964 Oct;24:1634–1643. [PubMed] [Google Scholar]
  12. KIM J. H., EIDINOFF M. L. ACTION OF 1-BETA-D-ARABINOFURANOSYLCYTOSINE ON THE NUCLEIC ACID METABOLISM AND VIABILITY OF HELA CELLS. Cancer Res. 1965 Jun;25:698–702. [PubMed] [Google Scholar]
  13. KONIGSBERG I. R. Clonal analysis of myogenesis. Science. 1963 Jun 21;140(3573):1273–1284. doi: 10.1126/science.140.3573.1273. [DOI] [PubMed] [Google Scholar]
  14. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  15. MOSKOWITZ M. GROWTH, DIFFERENTIATION AND REPRODUCTION OF AGGREGATES OF CULTURED MAMMALIAN CELLS. Nature. 1964 Sep 19;203:1233–1236. doi: 10.1038/2031233a0. [DOI] [PubMed] [Google Scholar]
  16. PUCK T. T., MARCUS P. I., CIECIURA S. J. Clonal growth of mammalian cells in vitro; growth characteristics of colonies from single HeLa cells with and without a feeder layer. J Exp Med. 1956 Feb 1;103(2):273–283. doi: 10.1084/jem.103.2.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. ROBBINS E., MARCUS P. I. MITOTICALLY SYNCHRONIZED MAMMALIAN CELLS: A SIMPLE METHOD FOR OBTAINING LARGE POPULATIONS. Science. 1964 May 29;144(3622):1152–1153. doi: 10.1126/science.144.3622.1152. [DOI] [PubMed] [Google Scholar]
  18. Silagi S. Hybridization of a malignant melanoma cell line with L cells in vitro. Cancer Res. 1967 Nov;27(11):1953–1960. [PubMed] [Google Scholar]
  19. Silagi S. Metabolism of 1-beta-D-arabinofuranosylcytosine in L cells. Cancer Res. 1965 Oct;25(9):1446–1453. [PubMed] [Google Scholar]
  20. Stubblefield E., Klevecz R. Synchronization of Chinese hamster cells by reversal of colcemid inhibition. Exp Cell Res. 1965 Dec;40(3):660–664. doi: 10.1016/0014-4827(65)90244-2. [DOI] [PubMed] [Google Scholar]
  21. TERASIMA T., TOLMACH L. J. Growth and nucleic acid synthesis in synchronously dividing populations of HeLa cells. Exp Cell Res. 1963 Apr;30:344–362. doi: 10.1016/0014-4827(63)90306-9. [DOI] [PubMed] [Google Scholar]
  22. WHITTAKER J. R. CHANGES IN MELANOGENESIS DURING THE DEDIFFERENTIATION OF CHICK RETINAL PIGMENT CELLS IN CELL CULTURE. Dev Biol. 1963 Aug;8:99–127. doi: 10.1016/0012-1606(63)90028-9. [DOI] [PubMed] [Google Scholar]
  23. Young R. S., Fischer G. A. The action of arabinosylcytosine on synchronously growing populations of mammalian cells. Biochem Biophys Res Commun. 1968 Jul 11;32(1):23–29. doi: 10.1016/0006-291x(68)90420-8. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES