Development of an in vitro assay for high-throughput screening investigating the role of mesenchymal stem cells on castration resistant prostate cancer cell growth
Williamson, Alexander James
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Androgen deprivation therapy (ADT) can increase survival from prostate cancer by up to 2-3 years, but tumours invariably relapse into an ADT-unresponsive, incurable form, known as castrate resistant prostate cancer (CRPC). CRPC is more aggressive and more likely to metastasise to bone, worsening morbidity and mortality. Mesenchymal stem cells have been implicated in alteration of androgen signalling within prostate cancer cells and stimulation of metastasis and resistance to anti-tumour therapy, and thus may play an important role in the development of castration resistance. A high throughput screen to identify compounds that inhibit the effect of MSCs on castration resistance would thus be valuable in development of novel chemotherapeutics against CRPC. Clones of the human CWR22PC and murine Myc-CaP Bo prostate cancer cell lines were characterised by their reduced growth in response to androgen deprivation, modelled using charcoal stripped serum and the antiandrogen enzalutamide. Investigations were performed to optimise the miniaturisation of this assay. The effect of conditioned media from human or murine mesenchymal stem cells on this cell growth was then examined in the presence of androgen and androgen deprivation, in a high-throughput format. It was found that MSC-conditioned media had only a small positive effect stimulating growth in CWR22PC cells, greatest in the enzalutamide-treated condition. In the murine Myc-CaP Bo cell line clone 5GSH-6943#5, MSC-conditioned media significantly stimulated castration-resistant growth in the androgen deprivation condition but not in the presence of androgen. However, assay validation indicated that the assay developed for either cell line was not suitable for high-throughput drug screening in its current form. Further optimisation is thus required for use of the assays developed as a platform for high-throughput screening to investigate the effects of various therapeutic compounds on MSC stimulation of castration-resistant prostate cancer cell growth.