Prostate specific antigen and androgen receptor induction and characterization of an immortalized adult human prostatic epithelial cell line

Abstract

Progress in prostate cancer research has been hindered by the lack of well characterized, immortalized, human prostatic epithelial cell lines that express markers of normal prostatic epithelial cells and mimic normal growth and differentiation responses to androgens. The objectives of this study were to: (i) establish immortalized cell lines from non-neoplastic, adult human prostatic epithelium using adenovirus-12/simian virus-40 (Ad12-SV40) hybrid virus; (ii) establish their prostatic epithelial origin; (iii) demonstrate androgen responsiveness; and (iv) examine response to growth factors. Primary epithelial cell cultures derived from a non-neoplastic, adult human prostate were infected with the Ad12-SV40 virus. Several immortalized clones were isolated. Single cell cloning of one clone, free of cytopathic effects, gave rise to the PWr-1E cell line. An immortalized cell line PWR-1E, which expresses many characteristics of normal prostatic epithelial cells was established. Immunostaining showed that cells express cytokeratins 8 and 18 normally expressed by differentiated, secretory prostatic epithelial cells. The most remarkable characteristics of PWR-1E cells are growth stimulation, increased expression of androgen receptor and induction of prostate specific antigen (PSA) expression in response to androgens, which indisputably establish their prostatic epithelial origin. They are positive for SV40 large-T antigen and show strong nuclear staining for p53. Cells from passages 23 and 40 were not tumorigenic in nude mice even when co-injected with Matrigel. They grow in a serum-free defined medium and respond to EGF, bFGF and TGF-beta. Passage 42-cells showed a human male (XY), hyperdiploid karyotype. The PWR-1E cell line is the only known Ad12-SV40-immortalized human prostatic epithelial cell line. PWR-1E cells can be used to study (i) the etiology and the multistep process of carcinogenesis and tumor progression in the human prostate; (ii) normal prostate physiology and differentiation; and (iii) potential prostate cancer chemopreventive agents.

Metastatic sublines of an SV40 large T antigen immortalized human prostate epithelial cell line

“…Most of the human prostate epithelial cells immortalized by viral transforming genes either lack androgen receptor expression or fail to respond to androgens, with the exception of PWR-1E [20].…”

“…Several laboratories have used either SV40 or HPV viral DNA to immortalize human non-neoplastic prostate epithelial cells, thereby creating valuable resources [13][14][15][16][17][18][19][20]. None of these lines is tumorigenic in athymic nude mice unless a second modification is introduced, such as transfection of K-ras [17] or exposure to radiation [21].…”

Hyaluronan stimulates transformation of androgen-independent prostate cancer

“…For these reasons, studies of HA-CD168 interaction were carried out primarily in PC3 cells. The PC3 cell line was isolated from a bone metastasis of HRCaP, which may have preserved more HA-responding features than the other two cell lines derived from metastatic brain and lymph nodes, respectively (30).…”

Immortalized and tumorigenic adult human prostatic epithelial cell lines: Characteristics and applications part I. Cell markers and immortalized nontumorigenic cell lines

“…Media developed for human keratinocyte growth, such as keratinocyte serum-free medium (K-SFM, Gibco, Grand Island, NY) and keratinocyte growth medium (KGM, Clonetics, San Diego, CA) have been found to be useful for growing and maintaining human prostatic epithelial cells [12,13]. The use of human cells is highly desirable for in vitro studies on human cancers.…”

“…The reasons is that other cell types are also present in prostatic tissue and include fibroblasts, smooth muscle, and endothelial cells. Epithelial origin is established by examining cytokeratin expression [12,13]. It is also necessary to establish that the cells are not of endothelial or muscle origin by demonstrating absence of factor VIII and desmin protein expression, respectively [ 12,131.…”