Detection of intracellular granularity induction in prostate cancer cell lines by small molecules using the HyperCyt high-throughput flow cytometry system

J Biomol Screen. 2009 Jul;14(6):596-609. doi: 10.1177/1087057109335671. Epub 2009 May 21.

Abstract

Prostate cancer is a leading cause of death among men due to the limited number of treatment strategies available for advanced disease. Discovery of effective chemotherapeutics involves the identification of agents that inhibit cancer cell growth. Increases in intracellular granularity have been observed during physiological processes that include senescence, apoptosis, and autophagy, making this phenotypic change a useful marker for identifying small molecules that induce cellular growth arrest or death. In this regard, epithelial-derived cancer cell lines appear uniquely susceptible to increased intracellular granularity following exposure to chemotherapeutics. We have established a novel flow cytometry approach that detects increases in side light scatter in response to morphological changes associated with intracellular granularity in the androgen-sensitive LNCaP and androgen-independent PC3 human prostate cancer cell lines. A cell-based assay was developed to screen for small molecule inducers of intracellular granularity using the HyperCyt high-throughput flow cytometry platform. Validation was performed using the Prestwick Chemical Library, where known modulators of LNCaP intracellular granularity, such as testosterone, were identified. Nonandrogenic inducers of granularity were also detected. A further screen of approximately 25,000 small molecules led to the identification of a class of aryl-oxazoles that increased intracellular granularity in both cell lines, often leading to cell death. The most potent agents exhibited submicromolar efficacy in LNCaP and PC3 cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Androgens / analysis
  • Androgens / pharmacology
  • Antineoplastic Agents / analysis
  • Antineoplastic Agents / chemistry
  • Antineoplastic Agents / pharmacology*
  • Cell Line, Tumor
  • Dose-Response Relationship, Drug
  • Drug Screening Assays, Antitumor
  • Flow Cytometry / methods*
  • Humans
  • Intracellular Space / drug effects*
  • Intracellular Space / pathology*
  • Male
  • Metribolone / pharmacology
  • National Institutes of Health (U.S.)
  • Prostatic Neoplasms / pathology*
  • Small Molecule Libraries / analysis
  • Small Molecule Libraries / chemistry
  • Small Molecule Libraries / pharmacology*
  • United States

Substances

  • Androgens
  • Antineoplastic Agents
  • Small Molecule Libraries
  • Metribolone