Establishment of an in vitro cell model system to study human prostate carcinogenesis.
TL;DR: A consistent change in chromosome 5 is observed in an in vitro cell model of human prostate carcinogenesis in which the near-diploid cells from the surrounding tissue of an adenocarcinoma of the prostate obtained from a 42-year-old patient were subjected to in vitrocell culture and passages.
Abstract: A positive family history of prostate cancer is a risk factor for this disease, suggesting that alterations of certain genes may play an important role in the development and progression of prostate cancer. However, genetic alterations responsible for initiation and acquisition of metastatic phenotypes by prostate cancer are not well defined. We have observed a consistent change in chromosome 5 in an in vitro cell model of human prostate carcinogenesis in which the near-diploid cells from the surrounding tissue of an adenocarcinoma of the prostate obtained from a 42-year-old patient were subjected to in vitro cell culture and passages. We have examined three different passages of this cell strain by conventional and molecular cytogenetic methods and have seen an increased number of alterations in chromosome 5 in higher passage cells, with accompanying changes in cell morphology. In late passages of this cell line, no cell showed two normal copies of chromosome 5 as analyzed by G-banding and fluorecent in situ hybridization (FISH). The long arm (q) of chromosome 5 was either missing or involved in structural rearrangements. This observation suggests that the q arm of chromosome 5 may carry a tumor suppressor gene(s) that is well-expressed in normal prostate tissue, but when one of these tumor suppressor gene(s) is mutated or deleted and its encoded mRNA and protein are differentially expressed or not expressed at all in the prostate cells, then it may lead to initiation of tumor growth and development. Cytogenetic analyses of early passage cells in this cell strain revealed that approximately 78.8% of metaphases were normal, with a 46,XY chromosome constitution, and 21.2% of cells had clonal alterations mostly of chromosomes 5, 7, 8, 15, 16 and Y. In the middle passages, abnormal cells increased in number (78.26%) and also showed a large number of chromosomal changes. In the late passages, all cells showed structural and numerical abnormalities of the same chromosomes, in addition to some new markers; no cells were found to have a normal karyotype. These chromosomal aberrations could be considered early markers of prostate carcinogenesis. Some of the markers present in late passage cells were similar to those reported in a well-characterized prostate cancer cell line, LNCaP.
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TL;DR: The hypothesis that resembles “adaptive mutation” such as has been described in bacteria subjected to nutritional starvation is supported, and appears to be driven by specific microenvironmental factors surrounding the tumor cell epithelium grown as 3D prostate organoids.
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 organo...
83 citations
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...In further investigations, we utilized a three-dimensional (3D) growth model, the rotating-wall vessel (RWV), to culture LNCaP cells either alone on the microcarrier beads, or in close contact with either prostate (Ozen et al., 1996) or bone stromal ceils that have been coated previously onto microcarrier beads in the RWV system (Zhau et al....
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...Human prostate stromal cells were derived from prostatectomy specimens obtained from men subjected to prostate cancer surgery; prostate stromal cells were derived from prostate tissues harvested from the contralateral side of a prostate cancer specimen, as described previously (Ozen et al., 1996)....
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...1 X ]06 cells resuspended in 25 txl of T media containing 5% FBS were injected orthotopically to the dorsolateral lobe of the prostate gland in castrated athymic mice, to assess their tumorigenic and metastatic potential in vivo (Gleave et al., 1992; Thalmann et al., 1994; Ozen et al., 1996; Zhau et al., 1997)....
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Journal Article•
TL;DR: Although few shifts in integrin expression were found to accompany disease progression, integrin heterodimer usage did change significantly, and the more metastatic sublines were distinct in their use of alphavbeta3 and, when compared with parental LNCaP cells, showed a shift in alpha6heterodimerization, a subunit critical not only for interaction with prostate basal lamina but also with the bone matrix, a favored site of prostate cancer metastases.
Abstract: During prostate cancer progression, invasive glandular epithelial cells move out of the ductal-acinar architecture and through the surrounding basement membrane. Extracellular matrix proteins and associated soluble factors in the basal lamina and underlying stroma are known to be important regulators of prostate cell behaviors in both normal and malignant tissues. In this study, we assessed cell interactions with extracellular matrix and stromal factors during disease progression by characterizing integrin usage and expression in a series of parental and lineage-derived LNCaP human prostate cancer cell lines. Although few shifts in integrin expression were found to accompany disease progression, integrin heterodimer usage did change significantly. The more metastatic sublines were distinct in their use of avb3 and, when compared with parental LNCaP cells, showed a shift in a6 heterodimerization, a subunit critical not only for interaction with prostate basal lamina but also for interaction with the bone matrix, a favored site of prostate cancer metastases.
76 citations
TL;DR: It is proposed that cell surface localized nucleolin protein may function in these cells as a novel HGF receptor and implicated the protein nucleolin in HGF signal reception by the LNCaP model prostate cancer cells.
Abstract: Background
Prostate cancer cells communicate reciprocally with the stromal cells surrounding them, inside the prostate, and after metastasis, within the bone. Each tissue secretes factors for interpretation by the other. One stromally-derived factor, Hepatocyte Growth Factor (HGF), was found twenty years ago to regulate invasion and growth of carcinoma cells. Working with the LNCaP prostate cancer progression model, we found that these cells could respond to HGF stimulation, even in the absence of Met, the only known HGF receptor. The new HGF binding partner we find on the cell surface may help to clarify conflicts in the past literature about Met expression and HGF response in cancer cells.
72 citations
Journal Article•
TL;DR: The results show that IGFBP-rP1 is expressed in both in situ and invasive prostate neoplasms, but not typically in normal secretory or BPH epithelium, and can be induced in human prostate cancer cell lines in vivo on interaction with an appropriate host environment.
Abstract: Many of the alterations in the insulin-like growth factor (IGF) axis in prostatic disease have been associated with changes in the insulin-like growth factor binding proteins (IGFBPs), a multigene family of proteins that are thought to mediate the action of IGFs on target tissues. IGFBP-related protein 1 (rP1), also known as IGFBP-7 or mac25, is a recently described member of the IGFBP family, the biological function of which has yet to be completely ascertained. In this study, we analyzed the localization of IGFBP-rP1 in prostate cancer and benign prostate tissues using immunohistochemistry and a polyclonal antibody, T1A12, that is specific for IGFBP-rP1. The most intense staining was observed in nerves, whereas smooth muscle cells in the prostate stained weakly. Lymphocytes were always negative. When normal prostatic secretory epithelium was present, staining was usually absent. The lining secretory epithelium stained positively in 0 of 12 (0%) cases of benign prostatic hyperplasia, 57 of 63 (90.5%) primary adenocarcinomas, and 7 of 7 (100%) prostate cancer metastases. Prostatic intraepithelial neoplasia showed a similar pattern of staining to that observed for the invasive tumors. Analysis of Northern blots showed that none of the prostate cancer cell lines (LNCaP, C4, C4-2, C4-2B4, 9069E3, DU145, and PC3) expressed IGFBP-rP1 mRNA. This lack of expression was confirmed by immunohistochemistry of s.c.-generated tumor xenografts of LNCaP and C4-2 and by immunoblot on serum-free-conditioned media from all prostatic cell lines. In contrast to these results, tumor xenografts generated by direct intraosseous injection of LNCaP or C4-2 to bone marrow space resulted in tumors that stained positively for IGFBP-rP1. Our results show that IGFBP-rP1 is expressed in both in situ and invasive prostate neoplasms, but not typically in normal secretory or BPH epithelium; furthermore, the expression of IGFBP-rP1 can be induced in human prostate cancer cell lines in vivo on interaction with an appropriate host environment.
43 citations
TL;DR: Because, in most cases, alteration of chromosome 5 resulted in the partial or complete loss of 5q, it was conjectured that 5q might contain one or more tumor-suppressor genes for human prostate cancer development.
21 citations