Affordable Access

Publisher Website

Intracellular trafficking as a determinant of AS-DACA cytotoxicity in rhabdomyosarcoma cells

Authors
Journal
BMC Cell Biology
1471-2121
Publisher
Springer (Biomed Central Ltd.)
Publication Date
Volume
12
Issue
1
Identifiers
DOI: 10.1186/1471-2121-12-36
Keywords
  • Research Article
Disciplines
  • Biology
  • Medicine

Abstract

Background Rhabdomyosarcoma (RMS) is a malignant soft tissue sarcoma derived from skeletal muscle precursor cells, which accounts for 5-8% of all childhood malignancies. Disseminated RMS represents a major clinical obstacle, and the need for better treatment strategies for the clinically aggressive alveolar RMS subtype is particularly apparent. Previously, we have shown that the acridine-4-carboxamide derivative AS-DACA, a known topoisomerase II poison, is potently cytotoxic in the alveolar RMS cell line RH30, but is 190-fold less active in the embryonal RMS cell line RD. Here, we investigate the basis for this selectivity, and demonstrate in these RMS lines, and in an AS-DACA- resistant subclone of RH30, that AS-DACA-induced cytotoxicity correlates with the induction of DNA double strand breaks. Results We show that inhibition of the multidrug-resistance associated protein (MRP1) has no effect on AS-DACA sensitivity. By exploiting the pH-dependent fluorescence properties of AS-DACA, we have characterized its intracellular distribution, and show that it concentrates in the cell nucleus, as well as in acidic vesicles of the membrane trafficking system. We show that fluorescence microscopy can be used to determine the localization of AS-DACA to the nuclear and cytoplasmic compartments of RMS cells grown as spheroids, penetrance being much greater in RH30 than RD spheroids, and that the vesicular signal leads the way into the spheroid mass. EEA1 and Rab5 proteins, molecular markers expressed on early-endosomal vesicles, are reduced by > 50% in the sensitive cell lines. Conclusion Taking the evidence as a whole, suggests that endosomal vesicle trafficking influences the toxicity of AS-DACA in RMS cells.

There are no comments yet on this publication. Be the first to share your thoughts.

Statistics

Seen <100 times
0 Comments

More articles like this

Sequestration of AS-DACA into Acidic Compartments...

on International Journal of Molec... Jun 25, 2013

Determinants of AQP6 trafficking to intracellular...

on Biology of the cell / under th... February 2006

W-7, a calmodulin inhibitor, potentiates dacarbazi...

on International Journal of Cance... May 15, 1987
More articles like this..