Androgen responsive adult human prostatic epithelial cell lines immortalized by human papillomavirus 18

Abstract

Prostate cancer and benign tumors of the prostate are the two most common neoplastic diseases in men in the United States, however, research on their causes and treatment has been slow because of the difficulty in obtaining fresh samples of human tissue and a lack of well characterized cell lines which exhibit growth and differentiation characteristics of normal prostatic epithelium. Non-neoplastic adult human prostatic epithelial cells from a white male donor were immortalized with human papillomavirus 18 which resulted in the establishment of the RWPE-1 cell line. Cells from the RWPE-1 cell line were further transformed by v-Ki-ras to establish the RWPE-2 cell line. The objectives of this study were to: (1) establish the prostatic epithelial origin and androgen responsiveness of RWPE-1 and RWPE-2 cell lines; (2) examine their response to growth factors; and (3) establish the malignant characteristics of the RWPE-2 cell line. Immunoperoxidase staining showed that both RWPE-1 and RWPE-2 cells express cytokeratins 8 and 18, which are characteristic of luminal prostatic epithelial cells, but they also coexpress basal cell cytokeratins. These cell lines show growth stimulation and prostate specific antigen (PSA) and androgen receptor (AR) expression in response to the synthetic androgen mibolerone, which establishes their prostatic epithelial origin. Both cell lines also show a dose-dependent growth stimulation by EGF and bFGF and growth inhibition when exposed to TGF-beta, however, the transformed RWPE-2 cells are less responsive. RWPE-1 cells neither grow in agar nor form tumors when injected into nude mice with or without Matrigel. However, RWPE-2 cells form colonies in agar and tumors in nude mice. In the in vitro invasion assay, RWPE-1 cells are not invasive whereas RWPE-2 cells are invasive. Nuclear expression of p53 and Rb proteins was heterogeneous but detectable by immunostaining in both cell lines. The RWPE-1 cells, which show many normal cell characteristics, and the malignant RWPE-2 cells, provide useful cell culture models for studies on prostate growth regulation and carcinogenesis.

Ornithine Decarboxylase Is Sufficient for Prostate Tumorigenesis via Androgen Receptor Signaling

“…This malignant phenotype was subsequently tested by s.c. injecting 1 Â 10 6 cells in 24 nonobese diabetic severe combined immunodeficiency g mice and monitoring them over a 6-month period. As previously described by the group that generated this normal prostate line, RWPE1 cells were nontumorigenic, 20 whereas tumor was palpated in 94% (45/48) of mice after injection with RWPE1_ODC cells ( Figure 2G). The mean time to palpation was 48.0 days (range, 29.0 to 81.0 days).…”

“…RWPE1 cells were generated by immortalization with HPV16 in 1997 by Bello et al 20 and characterized as nontumorigenic and noninvasive. The metastatic prostate cancer cells CWR22, LNCaP, and VCaP were purchased from ATCC and maintained in RPMI 1640 medium and Dulbecco's modified Eagle's medium with 10% fetal bovine serum at 37 C in a humidified air atmosphere at 5% CO 2 .…”

Selenoprotein expression is regulated at multiple levels in prostate cells

“…Surprisingly, a thorough examination of the effects of selenium supplementation on selenoprotein expression in prostate cells is lacking. In this study, we used the three common dietary supplements, sodium selenite, sodium selenate and selenomethionine (SeMet), to supplement the growth medium for "normal" prostate epithelial cells (RWPE-1; cells derived from normal prostate epithelia that were immortalized with human papolloma virus [17]) as well as androgen sensitive (LNCaP) and androgen insensitive (PC-3) prostate cancer cell lines. Because of their potential role in preventing transformation caused by DNA damage, we have chosen to focus on the three major intracellular antioxidant selenoproteins found in epithelial cells, GPX1, GPX2 and GPX4.…”

Rb/E2F4 and Smad2/3 link survivin to TGF-β-induced apoptosis and tumor progression

“…Both of them possess high levels of endogenous survivin, which was not suppressed by TGF-b1 (Figure 7b) even up to 72 h treatment (data not shown). Conversely, TGF-b downregulated survivin levels in RWPE-1 (Figure 7c), a non-tumorigenic human prostate epithelial cell line (Bello et al, 1997). We compared survivin levels in the context of tumor progression, using LNCaP cell line and its bone metastatic/androgen refractory variant, C4-2B, which express more survivin than LNCaP cells (Figure 7d).…”

“…LNCaP and C4-2B cells were maintained and treated as described for LNCaP (Chipuk et al, 2002). RWPE-1 cells (Bello et al, 1997) were cultured in complete keratinocyte serum-free growth medium (Invitrogen, Carlsbad, CA, USA). (GM2.1 medium: DMEM/F-12 supplemented with 5% FBS, 5 mg/ml insulin and 0.1 mM dexamethasone; GM3 medium: DMEM/F12 containing 1% FBS, 15 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) and 0.1 mM dexamethasone).…”