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. 2002 May;39(1):15–25. doi: 10.1023/A:1022435829894

Intracellular localisation studies of doxorubicin and Victoria Blue BO in EMT6-S and EMT6-R cells using confocal microscopy

SM Burrow 1, DA Phoenix 1, M Wainwright 1, MJ Tobin 2
PMCID: PMC3449804  PMID: 19003300

Abstract

The subcellular localisation of doxorubicin and Victoria Blue BO (VBBO) in a murine mammary tumour cell line EMT6-S, and the resistant sub-lineEMT6-R was studied, using confocal microscopy, in order to investigate their sites of action. In cells treated with doxorubicin (10 μ M) for 90 min, the pattern of intracellular drug distribution differed between the two cell lines. Doxorubicin was found to localise mainly in the nucleus of the sensitive cell line, whereas weak fluorescence was observed in the cytoplasm of the resistant cells, in a punctuate pattern, with no nuclear involvement. The drug also appeared to be effluxed more rapidly by the resistant cell line. The accumulation of doxorubicin at various time intervals over 1h in EMT6-S cells showed that the drug clearly interacted with both the plasma membrane and the nucleus. In contrast to doxorubicin, the intracellular distribution of VBBO in both EMT6-S and EMT6-R was similar, VBBO was clearly localised throughout the cytoplasm, in a punctuate pattern, which may be consistent with the widespread distribution of mitochondria. A more apical pattern of accumulation was noted in the EMT6-R cell line. No interaction with the plasma membrane was evident. These results indicate that the main modes of action for the two drugs differ markedly, suggesting involvement of both the membrane and the nucleus in the case of doxorubicin, but mitochondrial involvement for VBBO.

Keywords: Confocal, Doxorubicin, EMT6-R, EMT6-S, MDR, Victoria Blue BO

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