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

Abstract

Purpose: We have investigated if sequestration of metabolically dysfunctional mitochondria by theautophagosomes (mitophagy) imparts anti-androgen resistance. Method: Effects of the anti-androgen enzalutamide on the autophagy and mitophagy of androgen-dependentLNCaP and independent C4-2 cells are studied first. Autophagy is monitored by cellular fluorescence incells treated with monodansylcadavarine (MDC) or stained with anti-LC3B antibody. Cellular 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 fluorescence due 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 fluorescence due to Mitosox dye oxidation is used to identify mitochondria producing high superoxide (O2-). Mitophagyis monitored using fluorescence resonance energy transfer (FRET) by visualization of FRET images andquantitation 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 LNCaPcells than in independent C4-2 cells, both growing in androgen-depleted media. Enzalutamide treatmentinduces mitophagy in both cell lines, but the increase in mitophagy is more pronounced in the enzalutamideresistantC4-2 than in the sensitive LNCaP cells.

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Document Details

Document Type
Technical Report
Publication Date
Nov 01, 2017
Accession Number
AD1049173

Entities

People

  • George Wilding

Organizations

  • University of Texas at Austin

Tags

DTIC Thesaurus Topics

  • Androgens
  • Antibodies
  • Autophagy
  • Cell Line
  • Cells
  • Confocal Microscopy
  • Contracts
  • Data Analysis
  • Department Of Defense
  • Energy
  • Energy Transfer
  • Medical Personnel
  • Microscopes
  • Microscopy
  • Prostate
  • Prostate Cancer
  • Resistance

Fields of Study

  • Biology
  • Medicine

Readers

  • Chemistry (specifically Chemical Fluorescence)
  • Prostate Cancer Biology.