Summary
Resistance of leukemia cells tol-asparaginase is presumed to be due to increased expression of asparagine synthetase activity by resistant cells, so they are no longer dependent on an exogenous source ofl-asparagine for growth. The mechanism by which cells acquire the ability for increased enzyme expression, however, has not been clearly defined. Evidence presented here indicates that genomic alterations in the form of translocations, gene amplification, or increased P-glycoprotein expression, do not account for the phenotypic transformation froml-asparaginase sensitivity tol-asparaginase resistance. Instead, both sensitive and resistant L5178Y cells contain immunoreactive material detected by Western blotting with an antiserum prepared against bovine pancreatic asparagine synthetase. This suggests that the mechanism of resistance might involve modification of asparagine synthetase inl-asparaginase-resistant cells by an as-yet-unidentified mechanism or by inhibition of enzyme activity in thel-asparaginase-sensitive cells.
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Abbreviations
- CHO:
-
Chinese hamster ovary
- DMEM:
-
Dulbecco's modified Eagle medium
- EtBr:
-
ethidium bromide
- I.U.:
-
international unit
- PBS:
-
0.14 M NaCl, 0.01 M KCl, 0.02 M phosphate, pH 7.4
- SDS:
-
sodium dodecyl sulfate
References
Andrulis I, Chen J, Ray PN (1987) Isolation of human cDNAs for asparagine synthetase and expression in Jensen rat sarcoma cells. Mol Cell Biol 7: 2435–2443
Boyse EA, Old LJ, Campbell HA, Mashburn LT (1967) Suppression of murine leukemias byl-asparaginase. J Exp Med 125: 17–31
Bradford MM (1976) A rapid and sensitive method for the quantitation of micro quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72: 248–254
Colofiore J, Morrow J, Patterson M Jr (1973) Asparagine-requiring tumor cell lines and their non-requiring variants: cytogenetics, biochemistry, and population dynamics. Genetics 75: 503–514
Committee on Standardized Nomenclature for Mice (1972) Standard karyotype of the mouse,mus musculus. J Hered 63: 69–72
Cooney DA, Handschumachar RE (1970)l-asparaginase andl-asparagine metabolism. Ann Rev Pharmacol 10: 421–440
Fischer GA (1957) Studies of the culture of leukemia cells in vitro. Ann NY Acad Sci 76: 673–680
Fu S, Winchester R, Rai K, Kunkel H (1974) Hairy cell leukemia: proliferation of a cell with phagocytic and B-lymphocyte properties. Scand J Immunol 3: 847–851
Gerlach JH, Kartner N, Bell DR, Ling V (1986) Multidrug resistance. Cancer Surv 5: 25–46
Horowitz B, Madras BK, Old LJ, Boyce EJ, Meister A (1968) Asparagine synthetase activity of mouse leukemias. Science 160: 533–535
Juliano R, Ling V (1976) A surface glycoprotein modulating drug permeability in Chinese hamster ovary cell mutants. Biochim Biophys Acta 455: 152–162
Kartner R, Riordan JR, Ling V (1983) Cell surface P-glycoprotein associated with multidrug resistance in mammalian cell lines. Science 221: 1285–1288
Kartner R, Evernden-Porelle D, Bradley G, Ling V (1985) Detection of P-glycoprotein in multidrug-resistant cell lines by monoclonal antibodies. Nature 316: 820–823
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685
Mehlhaff PM, Luehr CA, Schuster SM (1985) Studies on the ammonia-dependent reaction of beef pancreatic asparagine synthetase. Biochemistry 24: 1104–1110
Meister A (1974) Asparagine synthetase. In: Boyer PD (ed) The enzymes, vol 10. Academic Press, New York, pp 561–580
Nesbit MN, Francke U (1973) A system of nomenclature for band patterns of mouse chromosomes. Chromosoma 41: 145–158
Patterson MK, Jr, Orr GR (1967)l-asparagine biosynthesis by nutritional variants of the Jensen sarcoma. Biochem Biophys Res Commun 26: 228–233
Pragar MD, Bachynsky N (1968a) Asparagine synthetase in normal and malignant tissues: correlation with tumor sensitivity to asparaginase. Arch Biochem Biophys 127: 645–654
Pragar MD, Bachynsky N (1968b) Asparagine synthetase in asparaginase-resistant and-susceptible mouse lymphomas. Biochem Biophys Res Commun 31: 43–47
Pfeiffer N, Mehlhaff P, Wylie D, Schuster S (1987) Topographical separation of the catalytic sites of asparagine synthetase explored with monoclonal antibodies. J Biol Chem 262: 11565–11570
Reed KC, Mann DA (1985) Rapid transfer of DNA from agarose gels to nylon membranes. Nucl Acids Res 3: 7207–7221
Schendl W (1971) The karyotype of the mouse. Chromosoma 35: 111–116
Schimke R, Brown P, Kaufman R, McGrogan M, Slate D (1981) Chromosomal and extrachomosomal localization of amplified dihydrofolate reductase genes in cultured mammalian cells. Cold Spring Harbor Symp Quant Biol 55: 785–797
Schirch L, Gross T (1968) Serine transhydroxymethylase. Identification as the threonine and allothreonine aldolases. J Biol Chem 243: 5651–5661
Sun NC, Chu EHY, Chang CC (1974) Staining method for the banding patterns of human mitotic chromosomes. Caryologia 27: 315–324
Tower DB, Peters EL, Curtis WC (1963) Guinea pigl-asparaginase. Properties, purification, and application to determination of asparagine in biological samples. J Biol Chem 238: 983–995
Unnithan S, Moraga A, Schuster SM (1984) A high-performance liquid chromatography assay for asparagine synthetase. Anal Biochem 136: 195–201
Yang TJ, Haddad-Khairallah L, Wachtel A (1981) Morphologic changes in the murine leukemia L5178Y cells treated with antibodies in the absence of complement activity. Exp Mol Pathol 35: 137–152
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Martin, J.K., Sun, W., Moraga-A, D. et al. An investigation into the mechanism ofl-asparaginase resistance in L5178Y murine leukemia cells. Amino Acids 5, 51–69 (1993). https://doi.org/10.1007/BF00806192
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DOI: https://doi.org/10.1007/BF00806192