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: Article

47 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 originalEnglish
Páginas (desde-hasta)1053-1061
Número de páginas9
PublicaciónCell Cycle
Volumen8
N.º7
DOI
EstadoPublished - 1 abr 2009

Huella dactilar

Prostate
Carrier Proteins
Adenosine Triphosphate
Population
Cell Line
Prostatic Neoplasms
Stem Cells
Side-Population Cells
DyeCycle Violet
Neoplastic Stem Cells
Fluorescent Dyes
Transgenic Mice
DNA Damage
Fluorescent Antibody Technique
Cultured Cells
Adenocarcinoma
Bone Marrow
Cell Membrane
Polymerase Chain Reaction
Membranes

ASJC Scopus subject areas

  • Molecular Biology
  • Developmental Biology
  • Cell Biology

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
Mathew, Grinu ; Timm, Earl A. ; Sotomayor, Paula ; Godoy, Alejandro ; Montecinos, Viviana P. ; Smith, Gary J. ; Huss, Wendy J. / ABCG2-mediated DyeCycle Violet efflux defined side population in benign and malignant prostate. En: Cell Cycle. 2009 ; Vol. 8, N.º 7. pp. 1053-1061.
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abstract = "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.",
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Mathew, G, Timm, EA, Sotomayor, P, Godoy, A, Montecinos, VP, Smith, GJ & Huss, WJ 2009, 'ABCG2-mediated DyeCycle Violet efflux defined side population in benign and malignant prostate', Cell Cycle, vol. 8, n.º 7, pp. 1053-1061. https://doi.org/10.4161/cc.8.7.8043

ABCG2-mediated DyeCycle Violet efflux defined side population in benign and malignant prostate. / Mathew, Grinu; Timm, Earl A.; Sotomayor, Paula; Godoy, Alejandro; Montecinos, Viviana P.; Smith, Gary J.; Huss, Wendy J.

En: Cell Cycle, Vol. 8, N.º 7, 01.04.2009, p. 1053-1061.

Resultado de la investigación: Article

TY - JOUR

T1 - ABCG2-mediated DyeCycle Violet efflux defined side population in benign and malignant prostate

AU - Mathew, Grinu

AU - Timm, Earl A.

AU - Sotomayor, Paula

AU - Godoy, Alejandro

AU - Montecinos, Viviana P.

AU - Smith, Gary J.

AU - Huss, Wendy J.

PY - 2009/4/1

Y1 - 2009/4/1

N2 - 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.

AB - 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.

KW - ABCG2

KW - DyeCycle Violet

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KW - Prostate cancer

KW - Side population

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Mathew G, Timm EA, Sotomayor P, Godoy A, Montecinos VP, Smith GJ y otros. ABCG2-mediated DyeCycle Violet efflux defined side population in benign and malignant prostate. Cell Cycle. 2009 abr 1;8(7):1053-1061. https://doi.org/10.4161/cc.8.7.8043