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Published on: November 13, 2015
Detection of intracellular granularity induction in prostate cancer cell lines by small molecules using the HyperCyt
Mark K Haynes1, J Jacob Strouse, Anna Waller
1University of New Mexico Center for Molecular Discovery, Albuquerque, New Mexico 87131, USA. MHaynes@salud.unm.edu
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.
Insights
Researchers identified a novel method to screen for prostate cancer drugs by detecting increased intracellular granularity. This approach led to the discovery of aryl-oxazole compounds that effectively reduce cancer cell growth and induce cell death.
Area of Science:
- Oncology
- Cell Biology
- Drug Discovery
Background:
- Prostate cancer remains a leading cause of male mortality, with limited therapeutic options for advanced stages.
- Identifying novel chemotherapeutics that inhibit cancer cell growth is crucial.
- Increased intracellular granularity is a marker for cellular processes like senescence, apoptosis, and autophagy, indicating growth arrest or death.
Purpose of the Study:
- To develop and validate a novel flow cytometry-based assay for screening small molecules that induce intracellular granularity in prostate cancer cells.
- To identify novel small molecules, particularly non-androgenic agents, that increase intracellular granularity and inhibit cancer cell growth.
Main Methods:
- Established a flow cytometry approach to detect increased side light scatter, indicative of intracellular granularity changes in LNCaP and PC3 prostate cancer cell lines.
- Developed a cell-based assay using the HyperCyt high-throughput platform for screening small molecules.
- Validated the assay using the Prestwick Chemical Library and screened approximately 25,000 small molecules.
Main Results:
- The assay successfully identified known modulators of intracellular granularity, such as testosterone, in LNCaP cells.
- A screen of 25,000 small molecules identified a class of aryl-oxazoles that increased intracellular granularity in both LNCaP and PC3 cells.
- The most potent aryl-oxazole compounds demonstrated submicromolar efficacy and induced cell death.
Conclusions:
- The developed flow cytometry assay is effective for identifying inducers of intracellular granularity in prostate cancer cells.
- Aryl-oxazoles represent a promising class of compounds for prostate cancer therapy, showing potent anti-cancer activity.
- This screening approach facilitates the discovery of novel chemotherapeutics for advanced prostate cancer.
