title: Detection of intracellular granularity induction in prostate cancer cell lines by small molecules using the HyperCyt high-throughput flow cytometry system.
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.
authors.affiliation: University of New Mexico Center for Molecular Discovery, Albuquerque, New Mexico 87131, USA. MHaynes@salud.unm.edu
authors.forename: Mark K
authors.initials: MK
authors.lastname: Haynes
authors.forename: J Jacob
authors.initials: JJ
authors.lastname: Strouse
authors.forename: Anna
authors.initials: A
authors.lastname: Waller
authors.forename: Andrei
authors.initials: A
authors.lastname: Leitao
authors.forename: Ramona F
authors.initials: RF
authors.lastname: Curpan
authors.forename: Cristian
authors.initials: C
authors.lastname: Bologa
authors.forename: Tudor I
authors.initials: TI
authors.lastname: Oprea
authors.forename: Eric R
authors.initials: ER
authors.lastname: Prossnitz
authors.forename: Bruce S
authors.initials: BS
authors.lastname: Edwards
authors.forename: Larry A
authors.initials: LA
authors.lastname: Sklar
authors.forename: Todd A
authors.initials: TA
authors.lastname: Thompson
mesh_terms: D000728:Androgens / Q000032:analysis / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000032:analysis / Q000737:chemistry / Q000494:pharmacology*; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D004354:Drug Screening Assays, Antitumor; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D042541:Intracellular Space / Q000187:drug effects* / Q000473:pathology*; D008297:Male; D015741:Metribolone / Q000494:pharmacology; D009316:National Institutes of Health (U.S.); D011471:Prostatic Neoplasms / Q000473:pathology*; D054852:Small Molecule Libraries / Q000032:analysis / Q000737:chemistry / Q000494:pharmacology*; D014481:United States
chemical_list: D000728:Androgens; D000970:Antineoplastic Agents; D054852:Small Molecule Libraries; D015741:Metribolone