PERMANENT PHENOTYPIC AND GENOTYPIC CHANGES OF PROSTATE CANCER CELLS CULTURED IN A THREE-DIMENSIONAL ROTATING-WALL VESSEL

• Published in: In vitro cellular & developmental biology. Animal 2001-03, Vol.37 (3), p.127-140
• Main Authors: RHEE, HONG W, ZHAU, HAIYEN E, PATHAK, SEN, MULTANI, ASHA S, PENNANEN, SARI, VISAKORPI, TAPIO, CHUNG, LELAND W. K
• Format: Article
• Language: English
• Subjects: anchorage-dependent and -independent growth; Androgens; Animals; Antigens; Beads; Bones; Cell culture; Cell Culture Techniques; Cell Division - drug effects; Cell growth; Cell interactions; Cell lines; chromosomal losses and gains; Chromosome Aberrations; Chromosome Banding; Coculture Techniques; Cultured cells; Cytogenetic Analysis; Cytogenetics; EGF, bFGF, KGF, HGF/SF, IGF-1 growth factors; Epithelial cells; Epithelium; Estradiol - pharmacology; Estrogens; Genotype; Growth factors; Growth rate; Growth Substances - pharmacology; Humans; Hybridization; Male; Metastases; Metribolone - pharmacology; Mice; Mice, Nude; microgravity-simulated cell culture; Microspheres; Mutation; NASA Biotechnology: Cell Science in Microgravity; Neoplasm Metastasis; Neoplasm Transplantation; Organoids; Phenotype; Prostate; Prostate cancer; prostate cancer cells in 3D culture; prostate cancer metastasis; prostate cancer models; prostate cancer–bone stroma interactions; Prostate-Specific Antigen; Prostatic Neoplasms - genetics; Prostatic Neoplasms - pathology; responsiveness to androgens and estrogens; responsiveness to growth factors; Rotation; SPECIAL - NASA/JOHNSON SPACE CENTER WORKSHOP; Stromal Cells; stromal–epithelial interactions; Tumor cell line; Tumor Cells, Cultured; Tumorigenicity; Tumors; Vessels
• ISSN: 1071-2690; 1543-706X; 1543-706X
• DOI: 10.1290/1071-2690(2001)037<0127:PPAGCO>2.0.CO;2

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.