Decreased drug accumulation in a mitoxantrone-resistant gastric carcinoma cell line in the absence of P-glycoprotein

Udo Kellner, Lesley Hutchinson, André Seidel, Hermann Lage, Mary K. Danks, Manfred Dietel, Scott H Kaufmann

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

An established gastric-carcinoma cell line, EPG85-257P, is extremely sensitive to mitoxantrone (IC50, 0.12 ng/ml). Stepwise selection with mitoxantrone for 3 years resulted in a cell line (EPG85-257RN) that is 7,056-fold resistant to mitoxantrone (IC50, 846 ng/ml) and displays cross-resistance to the topoisomerase(topo)-II poisons ametantrone (411x), etoposide(l 12x) and teniposide (60x) as well as the topo-I poisons 7-ethyl-10-hydroxycamptothecin (331x) and topotecan (58x). We now show that this resistance is multifactorial. Western blotting revealed a 5-fold decrease in topo-IIα polypeptide in the mitoxantrone-resistant cells. Immunohistochemistry and Western blotting failed to demonstrate F-glycoprotein overexpression. Formation of trapped topo-II-DNA complexes in the resistant cells required higher mitoxantrone concentrations than in parental cells, even though nuclei isolated from the EPG85-257RN cells formed cleavage complexes normally. In agreement with these observations, which suggest the possibility of a defect in mitoxantrone accumulation, examination of mitoxantrone accumulation in both cell lines by confocal laser microscopy revealed that the EPG85-257RN cells accumulate less mitoxantrone at steady state. From these results, we propose that mitoxantrone accumulation, along with alterations in topo-IIα expression, contribute to the resistance to mitoxantrone observed in these cells.

Original languageEnglish (US)
Pages (from-to)817-824
Number of pages8
JournalInternational Journal of Cancer
Volume71
Issue number5
DOIs
StatePublished - 1997

Fingerprint

Mitoxantrone
P-Glycoprotein
Stomach
Carcinoma
Cell Line
Type II DNA Topoisomerase
Pharmaceutical Preparations
irinotecan
Poisons
ametantrone
Confocal Microscopy
Inhibitory Concentration 50
Western Blotting
Teniposide
Topotecan
Type I DNA Topoisomerase
Etoposide
Cell Nucleus
Glycoproteins
Immunohistochemistry

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Decreased drug accumulation in a mitoxantrone-resistant gastric carcinoma cell line in the absence of P-glycoprotein. / Kellner, Udo; Hutchinson, Lesley; Seidel, André; Lage, Hermann; Danks, Mary K.; Dietel, Manfred; Kaufmann, Scott H.

In: International Journal of Cancer, Vol. 71, No. 5, 1997, p. 817-824.

Research output: Contribution to journalArticle

Kellner, Udo ; Hutchinson, Lesley ; Seidel, André ; Lage, Hermann ; Danks, Mary K. ; Dietel, Manfred ; Kaufmann, Scott H. / Decreased drug accumulation in a mitoxantrone-resistant gastric carcinoma cell line in the absence of P-glycoprotein. In: International Journal of Cancer. 1997 ; Vol. 71, No. 5. pp. 817-824.
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AU - Dietel, Manfred

AU - Kaufmann, Scott H

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