Logo of pnasPNASInfo for AuthorsSubscriptionsAboutThis Article
Proc Natl Acad Sci U S A. Jul 5, 1994; 91(14): 6674–6678.

Induction of differentiation in HL60 cells by the reduction of extrachromosomally amplified c-myc.


Oncogene amplification in tumor cells results in the overexpression of proteins that confer a growth advantage in vitro and in vivo. Amplified oncogenes can reside intrachromosomally, within homogeneously staining regions (HSRs), or extrachromosomally, within double minute chromosomes (DMs). Since previous studies have shown that low concentrations of hydroxyurea (HU) can eliminate DMs, we studied the use of HU as a gene-targeting agent in tumor cells containing extrachromosomally amplified oncogenes. In a neuroendocrine cell line (COLO 320), we have shown that HU can eliminate amplified copies of c-myc located on DMs, leading to a reduction in tumorigenicity in vitro and in vivo. To determine whether the observed reduction in tumorigenicity was due to differentiation, we next investigated whether HU could induce differentiation in HL60 cells containing extrachromosomally amplified c-myc. We compared the effects of HU, as well as two other known differentiating agents (dimethyl sulfoxide and retinoic acid), on c-myc gene copy number, c-myc expression, and differentiation in HL60 cells containing amplified c-myc genes either on DMs or HSRs. We discovered that HU and dimethyl sulfoxide reduced both c-myc gene copy number and expression and induced differentiation in cells containing c-myc amplified on DMs. These agents failed to have similar effects on HL60 cells with amplified c-myc in HSRs. By contrast, retinoic acid induced differentiation independent of the localization of amplified c-myc. These data illustrate the utility of targeting extrachromosomal DNA to modulate tumor phenotype and reveal that both HU and dimethyl sulfoxide induce differentiation in HL60 cells through DM elimination.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.3M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Snapka RM. Gene amplification as a target for cancer chemotherapy. Oncol Res. 1992;4(4-5):145–150. [PubMed]
  • Slamon DJ, Godolphin W, Jones LA, Holt JA, Wong SG, Keith DE, Levin WJ, Stuart SG, Udove J, Ullrich A, et al. Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. Science. 1989 May 12;244(4905):707–712. [PubMed]
  • Seeger RC, Brodeur GM, Sather H, Dalton A, Siegel SE, Wong KY, Hammond D. Association of multiple copies of the N-myc oncogene with rapid progression of neuroblastomas. N Engl J Med. 1985 Oct 31;313(18):1111–1116. [PubMed]
  • Johnson BE, Battey J, Linnoila I, Becker KL, Makuch RW, Snider RH, Carney DN, Minna JD. Changes in the phenotype of human small cell lung cancer cell lines after transfection and expression of the c-myc proto-oncogene. J Clin Invest. 1986 Aug;78(2):525–532. [PMC free article] [PubMed]
  • Carroll SM, DeRose ML, Gaudray P, Moore CM, Needham-Vandevanter DR, Von Hoff DD, Wahl GM. Double minute chromosomes can be produced from precursors derived from a chromosomal deletion. Mol Cell Biol. 1988 Apr;8(4):1525–1533. [PMC free article] [PubMed]
  • Von Hoff DD, Forseth B, Clare CN, Hansen KL, VanDevanter D. Double minutes arise from circular extrachromosomal DNA intermediates which integrate into chromosomal sites in human HL-60 leukemia cells. J Clin Invest. 1990 Jun;85(6):1887–1895. [PMC free article] [PubMed]
  • Von Hoff DD, Needham-VanDevanter DR, Yucel J, Windle BE, Wahl GM. Amplified human MYC oncogenes localized to replicating submicroscopic circular DNA molecules. Proc Natl Acad Sci U S A. 1988 Jul;85(13):4804–4808. [PMC free article] [PubMed]
  • Benner SE, Wahl GM, Von Hoff DD. Double minute chromosomes and homogeneously staining regions in tumors taken directly from patients versus in human tumor cell lines. Anticancer Drugs. 1991 Feb;2(1):11–25. [PubMed]
  • Gebhart E, Brüderlein S, Tulusan AH, von Maillot K, Birkmann J. Incidence of double minutes, cytogenetic equivalents of gene amplification, in human carcinoma cells. Int J Cancer. 1984 Sep 15;34(3):369–373. [PubMed]
  • Levan G, Ståhl F, Wettergren Y. Gene amplification in the murine SEWA system. Mutat Res. 1992 May;276(3):285–290. [PubMed]
  • Von Hoff DD, McGill JR, Forseth BJ, Davidson KK, Bradley TP, Van Devanter DR, Wahl GM. Elimination of extrachromosomally amplified MYC genes from human tumor cells reduces their tumorigenicity. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):8165–8169. [PMC free article] [PubMed]
  • Cahilly-Snyder L, Yang-Feng T, Francke U, George DL. Molecular analysis and chromosomal mapping of amplified genes isolated from a transformed mouse 3T3 cell line. Somat Cell Mol Genet. 1987 May;13(3):235–244. [PubMed]
  • Momand J, Zambetti GP, Olson DC, George D, Levine AJ. The mdm-2 oncogene product forms a complex with the p53 protein and inhibits p53-mediated transactivation. Cell. 1992 Jun 26;69(7):1237–1245. [PubMed]
  • Christen RD, Shalinsky DR, Howell SB. Enhancement of the loss of multiple drug resistance by hydroxyurea. Semin Oncol. 1992 Jun;19(3 Suppl 9):94–100. [PubMed]
  • Von Hoff DD, Waddelow T, Forseth B, Davidson K, Scott J, Wahl G. Hydroxyurea accelerates loss of extrachromosomally amplified genes from tumor cells. Cancer Res. 1991 Dec 1;51(23 Pt 1):6273–6279. [PubMed]
  • Snapka RM, Varshavsky A. Loss of unstably amplified dihydrofolate reductase genes from mouse cells is greatly accelerated by hydroxyurea. Proc Natl Acad Sci U S A. 1983 Dec;80(24):7533–7537. [PMC free article] [PubMed]
  • Von Hoff DD, Waddelow T, Forseth B, Davidson K, Scott J, Wahl G. Hydroxyurea accelerates loss of extrachromosomally amplified genes from tumor cells. Cancer Res. 1991 Dec 1;51(23 Pt 1):6273–6279. [PubMed]
  • Coppola JA, Cole MD. Constitutive c-myc oncogene expression blocks mouse erythroleukaemia cell differentiation but not commitment. Nature. 1986 Apr 24;320(6064):760–763. [PubMed]
  • DePinho RA, Schreiber-Agus N, Alt FW. myc family oncogenes in the development of normal and neoplastic cells. Adv Cancer Res. 1991;57:1–46. [PubMed]
  • Newburger PE, Chovaniec ME, Greenberger JS, Cohen HJ. Functional changes in human leukemic cell line HL-60. A model for myeloid differentiation. J Cell Biol. 1979 Aug;82(2):315–322. [PMC free article] [PubMed]
  • Pinkel D, Straume T, Gray JW. Cytogenetic analysis using quantitative, high-sensitivity, fluorescence hybridization. Proc Natl Acad Sci U S A. 1986 May;83(9):2934–2938. [PMC free article] [PubMed]
  • Carroll SM, DeRose ML, Gaudray P, Moore CM, Needham-Vandevanter DR, Von Hoff DD, Wahl GM. Double minute chromosomes can be produced from precursors derived from a chromosomal deletion. Mol Cell Biol. 1988 Apr;8(4):1525–1533. [PMC free article] [PubMed]
  • Von Hoff DD, Clark GM, Stogdill BJ, Sarosdy MF, O'Brien MT, Casper JT, Mattox DE, Page CP, Cruz AB, Sandbach JF. Prospective clinical trial of a human tumor cloning system. Cancer Res. 1983 Apr;43(4):1926–1931. [PubMed]
  • Dmitrovsky E, Kuehl WM, Hollis GF, Kirsch IR, Bender TP, Segal S. Expression of a transfected human c-myc oncogene inhibits differentiation of a mouse erythroleukaemia cell line. Nature. 1986 Aug 21;322(6081):748–750. [PubMed]
  • Prochownik EV, Kukowska J, Rodgers C. c-myc antisense transcripts accelerate differentiation and inhibit G1 progression in murine erythroleukemia cells. Mol Cell Biol. 1988 Sep;8(9):3683–3695. [PMC free article] [PubMed]
  • Holt JT, Redner RL, Nienhuis AW. An oligomer complementary to c-myc mRNA inhibits proliferation of HL-60 promyelocytic cells and induces differentiation. Mol Cell Biol. 1988 Feb;8(2):963–973. [PMC free article] [PubMed]
  • Lyman GH, Preisler HD, Papahadjopoulos D. Membrane action of DMSO and other chemical inducers of Friend leukaemic cell differentiation. Nature. 1976 Jul 29;262(5567):361–363. [PubMed]
  • Reboulleau CP, Shapiro HS. Chemical inducers of differentiation cause conformational changes in the chromatin and deoxyribonucleic acid of murine erythroleukemia cells. Biochemistry. 1983 Sep 13;22(19):4512–4517. [PubMed]
  • Heddle JA, Hite M, Kirkhart B, Mavournin K, MacGregor JT, Newell GW, Salamone MF. The induction of micronuclei as a measure of genotoxicity. A report of the U.S. Environmental Protection Agency Gene-Tox Program. Mutat Res. 1983 Sep;123(1):61–118. [PubMed]
  • Smith MA, Parkinson DR, Cheson BD, Friedman MA. Retinoids in cancer therapy. J Clin Oncol. 1992 May;10(5):839–864. [PubMed]
  • Glass CK, DiRenzo J, Kurokawa R, Han ZH. Regulation of gene expression by retinoic acid receptors. DNA Cell Biol. 1991 Nov;10(9):623–638. [PubMed]
  • Breitman TR, He RY. Combinations of retinoic acid with either sodium butyrate, dimethyl sulfoxide, or hexamethylene bisacetamide synergistically induce differentiation of the human myeloid leukemia cell line HL60. Cancer Res. 1990 Oct 1;50(19):6268–6273. [PubMed]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • Compound
    PubChem Compound links
  • MedGen
    Related information in MedGen
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...