Two sublines of the human T-lymphoblast leukemia cell line CCRF-CEM, which were resistant to methotrexate (MTX) due to defective MTX polyglutamate synthesis, were karyologically characterized. No statistically significant differences in the modal number of chromosomes were noted in resistant cells (CCRF-CEM/P) as compared to parent cells (91, range, 86-123; and 93, range; 78-103, respectively). Fifteen marker chromosomes were identified and their origins at least partially established. An isochromosome 7q, (marker 13) was present in all MTX-resistant cells but was not found in any sensitive cell karyotype. This marker chromosome may be involved in the emergence of drug-resistant cells from the parental population of CCRF-CEM cells. In all cell lines, chromosomes 8, 9 and 14 appear to be highly unstable and are involved in the genesis of many marker chromosomes. These chromosomes are also implicated in the in vivo genesis of various leukemias and lymphomas, which suggests that both in vivo tumor progression and in vitro cellular adaptation are marked by chromosome mutations that may activate multiple oncogenes.
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References
1.
AlamA., GeddesA.J., NorthA.C.T.: Methods and mechanisms for the development of MTX resistance in human lymphoid cells.Int. J. Cancer, 31: 45–50, 1983.
2.
BerensonR.J., FranckeU., DolnickB.J., BertinoJ.R.: Karyotypic analysis of methotrexate-resistant and sensitive mouse L5178Y cells.Cytogenet. Cell Genet., 29: 143–152, 1981.
3.
BertinoJ.R., SrimatkandadaS., CarmanM., JastreboffM., MelhmanL., MedinaW.D., MiniE.: Drug resistance: new approaches to treatment. In: New Experimental Modalities in the Control of Neoplasia.ChandraP. (Ed.), pp. 183–193, Plenum, New York, 1986.
4.
BrownP.C., BeverleyS.M., SchimkeR.T.: Relationship of amplified dihydrofolate reductase genes to double minute chromosomes in unstably resistant mouse fibroblast cell lines.Mol. Cell. Biol., 1: 1077–1083, 1981.
5.
CowanK.H., JolivetJ.: A methotrexate-resistant human breast cancer cell line with multiple defects including diminished formation of methotrexate polyglutamates.J. Biol. Chem., 259: 10793–10800, 1984.
6.
CurtG.A., CarneyD.N., CowanK.H., JolivetJ., BaileyB.D., DrakeJ.C., Kao-ShanC.S., MinnaJ.D., ChabnerB.A.: Unstable methotrexate resistance in human small-cell carcinoma associated with double minute chromosomes.N. Engl. J. Med., 308: 193–202, 1983.
7.
DolnickB.J., BerensonR.J., BertinoJ.R., KaufmanR.J., NunbergJ.H., SchimkeR.T.: Correlation of dihydrofolate reductase elevation with gene amplification in a homogeneously staining chromosomal region in L5178 cells.J. Cell Biol., 83: 394–402, 1979.
8.
EvaA., TronickS.R., GolR.A., PierceJ.H., AaronsonS.A.: Transforming genes of human hematopoietic tumors: frequent detection of ras-related oncogenes whose activation appears to be independent of tumor phenotype.Proc. Natl. Acad. Sci. USA, 80: 4926–4930, 1983.
9.
FoleyG.E., LazarusH., FarberS., UzmanB.G., BooneB.A., McCarthyR.E.: Continuous culture of human lymphoblasts from peripheral blood of a child with acute leukemia.Cancer, 18: 522–529, 1965.
10.
FreiE., RosowskyA., WrightJ.E., CucchiC.A., LippkeJ.A., ErvinT.J., JolivetJ., HaseltineW.A.: Development of methotrexate resistance in human squamous cell carcinoma of the head and neck in culture.Proc. Natl. Acad. Sci. USA, 81: 2873–2877, 1984.
11.
HasegawaT.: Inhibitory effects of ethidium bromide on mitotic chromosome condensation and its application to high-resolution chromosome banding.Cytogenet. Cell. Genet, 38: 56–61, 1984.
12.
JohnsD.G., BertinoJ.R.: Folate antagonists. In: Cancer Medicine.HollandJ.F., FreiE. (Eds.), pp. 775–790, Lea and Febiger, Philadelphia, 1982.
13.
JonesC., KaoF-T, TaylorR.T.: Chromosomal assignment of the gene for folylpolyglutamate synthetase to human chromosome 9.Cytogenet. Cell Genet, 28: 181–194, 1980.
14.
KaufmanR.J., SchimkeR.T.: Amplification and loss of dihydrofolate reductase genes in a Chinese hamster ovary cell line.Mol. Cell Biol., 1: 1069–1076, 1981.
15.
KaufmanR.J., BrownP.C., SchimkeR.T.: Amplified dihydrofolate reductase genes in unstably methotrexate resistant cells are associated with double minute chromosomes.Proc. Natl. Acad. Sci. USA, 76: 5669–5674, 1979.
16.
McBurneyM.W., WhitmoreG.F.: Isolation and biochemical characterization of folate deficient mutants of Chinese hamster cells.Cell, 2: 173–182, 1974.
17.
McCloskeyD.E., McGuireJ.J., RussellC.A., RowanB.G., BertinoJ.R., PizzornoG., MiniE.: Decreased polylpolyglutamate synthetase activity as a mechanism of methotrexate resistance in CCRF-CEM human leukemia sublines.J. Biol. Chem. (In press), 1991.
18.
MiniE., SrimatkandadaS., MedinaW.D., MorosonB.A., CarmanM.D., BertinoJ.R.: Molecular and karyological analysis of methotrexate-resistant and sensitive human leukemic CCRF-CEM cells.Cancer Res, 45: 317–324, 1985.
19.
NishimuraS., SekiyaT.: Human cancer and cellular oncogenes.Biochem. J., 243: 313–327, 1987.
20.
NunbergJ.H., KaufmanR.J., SchimkeR.T., UrlaubG., ChasinL.A.: Amplified dihydrofolate reductase genes are localized to a homogeneously staining region of a single chromosome in a methotrexate-resistant Chinese hamster ovary cell line.Proc. Natl. Acad. Sci., USA75: 5553–5556, 1978.
21.
PizzornoG., MiniE., CoronnelloM., McGuireJ.J., MorosonB., CashmoreA.R., DreyerR.N., LinJ.T., MazzeiT., PeritiP., BertinoJ.R.: Impaired polyglutamylation of methotrexate as a cause of resistance in CCRF-CEM cells after short-term, high-dose treatment with this drug.Cancer Res, 48: 2149–2155, 1988.
22.
SmallM.F., StanyonR., SmithD.G., SineoL.: High-resolution chromosomes of rhesus macaques (macaca mulatta).Am. J. Primatol, 9: 63–67, 1985.
23.
TrentJ.M., BuickR.N., OlsonS., HornsR.C.Jr., SchimkeR.T.: Cytologic evidence for gene amplification in methotrexate-resistant cells obtained from a patient with ovarian adenocarcinoma.J. Clin. Oncol, 2: 8–15, 1984.
24.
TsujimotoY., JaffeE., CossmanJ., GorhamJ., NowellP.C., CroceC.M.: Clustering of breakpoints on chromosome 11 in human B-cell neoplasm with the t(11; 14) chromosome translocation.Nature, 315: 340–343, 1985.
