Mitomycin C resistant L1210 leukemia cells

Association with pleiotropic drug resistance

Robert T Dorr, James D. Liddil, Jeffrey M. Trent, William S. Dalton

Research output: Contribution to journalArticle

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Abstract

A mitomycin C-resistant (MMCR) strain of L1210 mouse leukemia was developed by continuous drug exposure in vitro. MMC concentrations were increased in a stepwise fashion beginning at 0.033 μM and ending at 0.34 μM. This produced a 10-fold resistant cell line over the parental line. Resistance simultaneously developed to anthracene and anthracycline DNA intercalators, to vinca alkaloids and epipodophyllotoxins but not to cisplatin, bleomycin, fluorouracil or ionizing X-rays. MMC resistance was reversed using the membrane-active agent verapamil. The level of non-protein sulfhydryls was increased 2-fold in the MMCR cells. Intracellular uptake of unchanged MMC was reduced by 40% in the MMCR cells. Cytogenetic analyses demonstrated no recognizable clonal chromosomal alterations unique to the resistant subline and no evidence of double minutes or homogeneously staining regions in the DNA. Gel renaturation analysis failed to document the presence of an amplified DNA domain. Southern blotting of parental and MMCR DNA using a cDNA probe (CHP1) for the P-glycoprotein gene also failed to demonstrate amplification or rearrangement of P-glycoprotein-related homologous sequences. However, an Mr 180,000 glycoprotein was detected in the plasma membranes from MMCR cells. This protein also specifically reacted with a monoclonal antibody (C219) to the P-glycoprotein of Ling and co-workers [Kartner et al., Nature, Lond. 316, 820 (1985)]. These results suggest a pleiotropic drug resistance pattern in the MMCR cells, associated with membrane glycoprotein alterations, enhanced non-protein sulfhydryl levels, and reduced MMC accumulation. This is a novel observation for a resistant cell line selected with an alkylating agent.

Original languageEnglish (US)
Pages (from-to)3115-3120
Number of pages6
JournalBiochemical Pharmacology
Volume36
Issue number19
DOIs
StatePublished - Oct 1 1987

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Leukemia L1210
Mitomycin
Drug Resistance
P-Glycoprotein
Pharmaceutical Preparations
DNA
Cells
Podophyllotoxin
Vinca Alkaloids
Intercalating Agents
Cell Line
Cytogenetic Analysis
Alkylating Agents
Anthracyclines
Membrane Glycoproteins
Bleomycin
Cell membranes
Sequence Homology
Verapamil
Southern Blotting

ASJC Scopus subject areas

  • Pharmacology

Cite this

Mitomycin C resistant L1210 leukemia cells : Association with pleiotropic drug resistance. / Dorr, Robert T; Liddil, James D.; Trent, Jeffrey M.; Dalton, William S.

In: Biochemical Pharmacology, Vol. 36, No. 19, 01.10.1987, p. 3115-3120.

Research output: Contribution to journalArticle

Dorr, Robert T ; Liddil, James D. ; Trent, Jeffrey M. ; Dalton, William S. / Mitomycin C resistant L1210 leukemia cells : Association with pleiotropic drug resistance. In: Biochemical Pharmacology. 1987 ; Vol. 36, No. 19. pp. 3115-3120.
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abstract = "A mitomycin C-resistant (MMCR) strain of L1210 mouse leukemia was developed by continuous drug exposure in vitro. MMC concentrations were increased in a stepwise fashion beginning at 0.033 μM and ending at 0.34 μM. This produced a 10-fold resistant cell line over the parental line. Resistance simultaneously developed to anthracene and anthracycline DNA intercalators, to vinca alkaloids and epipodophyllotoxins but not to cisplatin, bleomycin, fluorouracil or ionizing X-rays. MMC resistance was reversed using the membrane-active agent verapamil. The level of non-protein sulfhydryls was increased 2-fold in the MMCR cells. Intracellular uptake of unchanged MMC was reduced by 40{\%} in the MMCR cells. Cytogenetic analyses demonstrated no recognizable clonal chromosomal alterations unique to the resistant subline and no evidence of double minutes or homogeneously staining regions in the DNA. Gel renaturation analysis failed to document the presence of an amplified DNA domain. Southern blotting of parental and MMCR DNA using a cDNA probe (CHP1) for the P-glycoprotein gene also failed to demonstrate amplification or rearrangement of P-glycoprotein-related homologous sequences. However, an Mr 180,000 glycoprotein was detected in the plasma membranes from MMCR cells. This protein also specifically reacted with a monoclonal antibody (C219) to the P-glycoprotein of Ling and co-workers [Kartner et al., Nature, Lond. 316, 820 (1985)]. These results suggest a pleiotropic drug resistance pattern in the MMCR cells, associated with membrane glycoprotein alterations, enhanced non-protein sulfhydryl levels, and reduced MMC accumulation. This is a novel observation for a resistant cell line selected with an alkylating agent.",
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