Abstract
A significant problem encountered in the treatment of cancer patients is that cancer cells often evolve resistance to chemotherapeutic agents. One of the mechanisms responsible for drug resistance is gene amplification. The study of the behavior of genes conferring drug resistance is very important to determine future treatments for cancer patients that will minimize the effect of gene amplification. One of the best methods to investigate this phenomenon is to use chromosome microdissection to directly access the amplified gene or genes. In the present study, chromosome microdissection and fluorescent in-situ hybridization (FISH) were applied for the identification of genes residing in a homogeneously staining region (HSR) in drug-resistant cell sublines developed by treatment of the T-cell leukemia cell line CCRF-CEM with increasing levels of the anthracycline, epirubicin. We have demonstrated that the selection by epirubicin actually elevated the level of the multidrug resistance-associated protein (MRP1) gene. We argue that the breakage fusion bridge (B-F-B) cycle offers a plausible explanation for this amplification. The DNA prepared from the amplified regions by chromosome microdissection provides a resource for future investigations looking for the possible presence of novel genes contributing to drug resistance.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Cherif D, Lavialle N, Modjtahedi M, Le Contait R, Berger, Brison O (1989) Selection of cells with different chromosomal localizations of the amplified c-myc gene during in vivo and in vitro growth of the breast carcinoma cell line SW 613-S. Chromosoma 97: 327–333.
Childs S, Ling V (1994) The MDR superfamily of genes and its biological implications. In Vincent T, Vita D, Hellman S, Rosenberg SA, eds., Important Advances in Oncology. Philadelphia: J. B Lippincott, pp. 21–36.
Cole SP, Bhardwaj G, Gerlach J et al. (1992) Overexpression of a transporter gene in a multidrug-resistant human lung cancer cell line. Science 258: 1650–1654.
Davey R, Longhurst T, Davey M et al. (1995) Drug resistance mechanisms and MRP expression in response to epirubicin treatment in a human leukaemia cell line. Leuk Res 19: 275–282.
Eijdems EW, De Hass M, Coco-Martin JM et al. (1995) Mechanisms of MRP over-expression in four human lung-cancer cell lins and analysis of the MRP amplicon. Int J Cancer 60: 676–684.
Filipits M, Suchomel RW, Zochbauer S, Brunner R, Lechner K, Pirker R (1997) Multidrug resistance-associated protein in acute myeloid leukemia: impact on treatment outcome. Clin Cancer Res 3: 1419–1425.
Filipits M, Stranzl T, Pohl G et al. (1999) MRP expression in acute myeloid leukemia. An update. Adv Exp Med Bio 457: 141–150.
Foley G, Lazarus H, Farber S, Uzman B, Boone B, Mc Carthy R (1965) Continuous culture of human lymphoblasts from peritoneal blood of a child with acute leukemia. Cancer 18: 522–529.
Grant CE, Valdimarson G, Hipfner DR, Almquist KC, Cole SP, Deeley RG (1994) Overexpression of multidrug resistance-associated protein (MRP1) increase resistance to natural product drugs. Cancer Res 54: 357–361.
Hunault M, Zhou D, Delmer A et al. (1997) Multidrug resistance gene expression in acute myeloid leukemia: Prognosis significance for in vivo drug resistance induction treatment. Ann Hematol 74: 65–71.
Kasimir-Bauer S, Ottinger H, Meusers P et al. (1998) In acute myeloid leukemia, coexpression of at least two proteins, including P-glycoprotein, the multidrug resistance-related protein, bcl-2, mutant p53, and heat-shock protein 27, is predictive of the response to induction chemotherapy. Exp Hematol 26: 1111–1117.
Kohler T, Leiblein S, Borchert S et al. (1999) Absolute levels of MDR-1, MRP, and BCL-2 MRNA and tumor remission in acute leukemia. Adv Exp Med Biol 457: 177–185.
Kool M, Linden M, Haas M, Baas F, Borst P (1999) Expression of human MRP16, a homologue of the multidrug resistance protein gene MRP11, in tissues and cancer cells. Cancer Res 59: 175–182.
Kuss BJ, O'Neill GM, Eyre H, Doggett NA, Callen DF, Davey RA (1998) ARA, a novel ABC transporter, is located at 16p13.1, is deleted in inv(16) leukemias, and is shown to be expressed in primitive hematopoietic precursors. Genomics 5: 455–458.
Lo AW, Sabatier L, Fouladi B, Pottier G, Ricoul M, Murnane JP (2002) DNA amplification by breakage/fusion/bridge cycles initiated by spontaneous telomere loss in a human cancer cell line. Neoplasia 4: 531–538.
Longhurst TJ, O'Neil GM, Harvie RM, Davey RA (1996) The anthracycline resistance-associated (ara) gene, a novel gene associated with multidrug resistance in a human leukaemia cell line. Br J Cancer 74: 1331–1335.
McClintock B (1941) Spontaneous alteration in chromosome size and form in Zea mays. Cold Spring Harbour Symp Quant Biol 9: 72–81.
McClintock B (1978) Mechanisms that rapidly reorganise the genome. Stadler Symp 10: 25–47.
McClintock B (1984) The significance of responses of the genome to challenge. Science 226: 792–801.
Obara K, Ghazizadeh M, Shimizu H et al. (2002) Comparative genomic hybridization study of genetic changes associated with vindesine resistance in esophageal carcinoma. Int J Oncol 20: 255–260.
Olinski R, Jaruga P, Foksinski M, Bialkowski K, Tujakowski J (1997) Epirubicin-induced oxidative DNA damage and evidence for its repair in lymphocytes of cancer patients who are undergoing chemotherapy. Mol Pharmacol 52: 882–885.
O'Neill GM, Peters GB, Harvie RM, Macknezie HB, Henness S, Davey RA (1998) Amplification and expression of the ABC transporters ARA and MRP1 in a series of multidrug-resistant leukaemia cell sub-lines. Br J Cancer 77: 2076–2080.
Reeve JC, Rabbitts PH, Twentyman PR (1989) Amplification and expression of mdr 1 gene in a multidrug resistant variant of small cell lung cancer cell line NCI-H69. Br J Cancer 60: 339–342.
Saunders RDC, Glover MD, Ashburner M et al. (1989) PCR amplification of DNA microdissected from a single polytene chromosome band: a comparison with conventional microcloning. Nucleic Acids Res 17: 9027–9037.
Schimke RT, Kaufman RJ, Alt FW, Kellems (1978) Gene amplification and drug resistance in cultures murine cells. Science 51: 455–458.
Slovak ML, Ho JP, Bhardwaj G, Kurz EU, Deeley RG, Cole SPC (1993) Localisation of a novel multidrug resistance associated gene in the HT 1080/DR4 and H69AR human tumour cell lines. Cancer Res 53: 3221–3225.
Smith KA, Gorman PA, Stark MB, Groves RP, Stark GR (1990) Distinctive chromosomal structures are formed very early in the amplification of CAD genes. Cell 63: 1219–1227.
Smith KA, Stark MB, Gorman PA, Stark GR (1992) Fusion near telomeres occur very early in the amplification of CAD genes in Syrian hamster cells. Proc Natl Acad Sci USA 89: 5427–5431.
Stark GR (1994) Regulation and mechanisms of mammalian gene amplification. Adv Cancer Res 61: 87–113.
Windle BE, Wahl GM (1992) Molecular dissection of mammalian gene amplification: new mechanistic insights revealed by analyses of very early events. Mutat Res 273: 199–224.
Zhou DC, Hoang-Ngoc L, Delmer A et al. (1994) Expression of resistance genes in acute leukemia. Leuk Lymph 13: 27–30.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Mahjoubi, F., Hill, R.J. & Peters, G.B. Chromosome microdissection identifies genomic amplifications associated with drug resistance in a leukemia cell line: an approach to understanding drug resistance in cancer. Chromosome Res 14, 263–276 (2006). https://doi.org/10.1007/s10577-006-1042-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10577-006-1042-9