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Autophagosomal Sequestration of Mitochondria as an Indicator of Antiandrogen Therapy Resistance of Prostate Cancer (PCa)

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Technical Report,01 Nov 2016,31 Oct 2017

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The University of Texas MD Anderson Cancer Center Houston United States

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Purpose We have investigated if sequestration of metabolically dysfunctional mitochondria by the autophagosomes mitophagy imparts anti-androgen resistance.Method Effects of the anti-androgen enzalutamide on the autophagy and mitophagy of androgen-dependent LNCaP and independent C4-2 cells are studied first. Autophagy is monitored by cellular fluorescence in cells treated with monodansylcadavarine MDC or stained with anti-LC3B antibody. Cellular fluorescencedue to Mitosox dye oxidation is used to identify mitochondria producing high superoxide O2-. Mitophagy is monitored using fluorescence resonance energy transfer FRET by visualization of FRET images and quantitation of FRET image intensities using a Nikon A1 or a Leica Di8 fluorescence confocal microscopeand Image J software. Results and Discussion Our data show that the degree of mitophagy is more in androgen-dependent LNCaP cells than in independent C4-2 cells, both growing in androgen-depleted media. Enzalutamide treatment induces mitophagy in both cell lines, but the increase in mitophagy is more pronounced in the enzalutamide-resistant C4-2 than in the sensitive LNCaP cells. Mitophagy in circulating tumor cells CTCs isolated from patient blood samples are currently being standardized.

Subject Categories:

  • Medicine and Medical Research
  • Biochemistry

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