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

Area of Science:

• Cell Biology
• Cancer Research
• Biochemistry

Background:

• ATP-binding cassette protein G2 (ABCG2) efflux identifies stem/progenitor cells as the side population (SP).
• Current methods for SP identification may require UV excitation, potentially damaging DNA.

Purpose of the Study:

• To evaluate DyeCycle Violet (DCV) as a novel vital dye for identifying ABCG2-mediated SP in prostate cells.
• To assess DCV's utility in isolating viable prostate stem/cancer stem cells.

Main Methods:

• Flow cytometry analysis of prostate cell lines and tissues using DCV.
• Inhibition of ABCG2 with Fumitremorgin C to confirm DCV efflux mechanism.
• qRT-PCR and immunofluorescence to confirm ABCG2 expression in SP cells.

Main Results:

• DCV efflux identified SPs in CWR-R1, DU-145, and RWPE-1 prostate cancer cell lines.
• SPs were also identified in TRAMP, human benign, and human prostate cancer tissues.
• Fumitremorgin C treatment abolished the DCV-identified SP, confirming ABCG2 mediation.

Conclusions:

• DCV is a viable substrate for ABCG2-mediated efflux, enabling SP identification in prostate cancer.
• DCV offers a UV-free method for isolating viable prostate stem/cancer stem cells.
• This method minimizes DNA damage risk in sorted cell populations.