Permanent Phenotypic and Genotypic Changes of Prostate Cancer Cells Cultured in a Three-Dimensional Rotating-Wall Vessel

Abstract

A three-dimensional (3D) integrated rotating-wall vessel cell-culture system was used to evaluate the interaction between a human prostate cancer cell line, LNCaP, and microcarrier beads alone, or microcarrier beads previously seeded with either prostate or bone stromal cells. Upon coculture of LNCaP cells with microcarrier beads either in the presence or in the absence of prostate or bone stromal cells, 3D prostate organoids were formed with the expected hormonal responsiveness to androgen, increased cell growth, and prostate-specific antigen production. In this communication, we define permanent phenotypic and genotypic changes of LNCaP cells upon coculture with microcarrier beads alone, or with microcarrier beads previously seeded with either prostate or bone stromal cells. Most notably, we observed selective genetic changes, i.e., chromosomal losses or gains, as evaluated by both conventional cytogenetic and comparative genomic hybridization, in LNCaP sublines derived from the prostate organoids. Moreover, the derivative LNCaP cells appear to have altered growth profiles, and exhibit permanent and stable changes in response to androgen, estrogen, and growth factors. The derivative LNCaP sublines showed increased anchorage-independent growth rate, and enhanced tumorigenicity and metastatic potential when inoculated orthotopically in castrated athymic mice. Our results support the hypothesis that further nonrandom genetic and phenotypic changes in prostate cancer epithelial cells can occur through an event that resembles "adaptive mutation" such as has been described in bacteria subjected to nutritional starvation. The occurrence of such permanent changes may be highly contact dependent, and appears to be driven by specific microenvironmental factors surrounding the tumor cell epithelium grown as 3D prostate organoids.

Immunohistochemical analysis of differentiation in static and mixed prostate cancer spheroids

“…In one study, the down-regulation in PSA expression correlated with the acquisition of androgen-independent cell growth [19]. The tissue was more tumorigenic and metastatic when implanted in castrated, athymic mice than the PSAexpressing control [20]. Consistent with these findings, a decrease in PSA expression within tumors is associated with higher incidence of fatalities in patients with prostate cancer [21].…”

Anti-metastatic effects of liposomal gemcitabine in a human orthotopic LNCaP prostate cancer xenograft model

“…During any of these steps, tumor cells interact with host factors in the microenvironment and will therefore will be subject to selection [57]. That there is actually a potential, selectable cellular heterogeneity also within PCa cell lines was shown by establishing LNCaP subclones displaying different tumor incidence or metastatic potential [58][59][60][61].…”

Microgravity assay of neuroblastoma: in vitro aggregation kinetics and organoid morphology correlate with MYCN expression

“…In particular, the lack of preferential orientation from gravitational force (Tsao et al 1991;Schwartz et al 1992), low shear , and absence of turbulence (Cherry and Papoutsakis 1986) promote spontaneous cell-cell interactions and aggregation and permit the formation and propagation of 3D structures, called organoids Hewitt et al 1999). In this environment, and within the organoid structure, a variety of cancer cells have been shown to better emulate their intrinsic in vivo biology, behavior, morphology, and humoral responses compared to traditional monolayer culture (Jessup et al 1993;Freed and Vunjak-Novakovic 1997;Ingram et al 1997;O'Connor 1999;Licato et al 2001;Rhee et al 2001;Vamvakidou et al 2007). …”