ABCG2-mediated DyeCycle Violet efflux defined side population in benign and malignant prostate

Grinu Mathew, Earl A. Timm, Paula Sotomayor, Alejandro Godoy, Viviana P. Montecinos, Gary J. Smith, Wendy J. Huss

Resultado de la investigación: Contribución a una revistaArtículo

48 Citas (Scopus)

Resumen

The efflux of Hoechst 33342 by ATP-binding cassette protein G2 (ABCG2) membrane pump allows reproducible identification of a subpopulation of cells by flow cytometric analysis termed the "side population" (SP). The SP identified by constitutive Hoechst efflux contains the stem/progenitor cell population from bone marrow and many solid organs, including prostate. DyeCycle Violet (DCV) is a cell membrane permeable, fluorescent vital dye that intercalates into DNA and is a substrate for ABCG2-mediated efflux. Therefore, DCV was evaluated in this study as a tool for identification of the SP from prostate cancer cell lines and from freshly harvested human prostate tissue. SPs that demonstrated ABCG2-mediated efflux of DCV were identified in the human prostate cancer cell lines CWR-R1, DU-145 and RWPE-1, but not in the BPH-1, LAPC-4 or PC-3 cell lines. Additionally, a SP was identified in enzymatically disaggregated prostate tumors from Transgenic Adenocarcinoma of Mouse Prostate (TRAMP), human benign prostate tissue and human prostate cancer tissue. The causal role of ABCG2-mediated efflux of DCV in the identification of the SP was confirmed by loss of the SP by incubation with the specific inhibitor of ABCG2, Fumitremorgin C. Expression of ABCG2 in the SP cells was confirmed by qRT-PCR and immunofluorescence analysis. Consequently, DCV represents an important new tool for isolation of viable candidate stem cells/cancer stem cells as a SP from cultured prostate cell lines, and prostate tissue specimens, without the requirement for instrumentation with ultra-violet excitation capability and minimizing the risk of damage to DNA in the sorted population.

Idioma originalInglés
Páginas (desde-hasta)1053-1061
Número de páginas9
PublicaciónCell Cycle
Volumen8
N.º7
DOI
EstadoPublicada - 1 abr 2009

Áreas temáticas de ASJC Scopus

  • Biología molecular
  • Biología del desarrollo
  • Biología celular

Huella Profundice en los temas de investigación de 'ABCG2-mediated DyeCycle Violet efflux defined side population in benign and malignant prostate'. En conjunto forman una huella única.

  • Citar esto

    Mathew, G., Timm, E. A., Sotomayor, P., Godoy, A., Montecinos, V. P., Smith, G. J., & Huss, W. J. (2009). ABCG2-mediated DyeCycle Violet efflux defined side population in benign and malignant prostate. Cell Cycle, 8(7), 1053-1061. https://doi.org/10.4161/cc.8.7.8043