cytokeratin 18, cytokeratin 8,xa0

Tumor Supressor Gene(s):xa0p53 +,xa0pRB + RefBello D, et al. Androgen responsive adult human prostatic epithelial cell lines immortalized by human papillomavirus 18. Carcinogenesis 18: 1215-1223, 1997. PubMed: 9214605

In 3-dimensional Matrigel culture, RWPE-1 cells organize into acini and secrete PSA into the lumen when exposed to androgen.xa0RefBello-DeOcampo D, et al. Laminin-1 and alpha6beta1 integrin regulate acinar morphogenesis of normal and malignant human prostate epithelial cells. Prostate 46: 142-153, 2001. PubMed: 11170142

When injected with Matrigel or with stromal cells, into male athymic rodents, RWPE-1 cells also organize into acini RefWebber MM, et al. Human cell lines as an in vitro/in vivo model for prostate carcinogenesis and progression. Prostate 47: 1-13, 2001. PubMed: 11304724 and produce PSA.xa0

Cells from the RWPE-1 cell line were further transformed by Ki-ras using the Kirstin murine sarcoma virus (Ki-MuSV) to establish the tumorigenic RWPE-2 cell line (ATCC CRL-11610) RefBello D, et al. Androgen responsive adult human prostatic epithelial cell lines immortalized by human papillomavirus 18. Carcinogenesis 18: 1215-1223, 1997. PubMed: 9214605 and the RWPE2-W99 (ATCC CRL-2853) cell line.xa0

Further, a family of tumorigenic cell lines, that mimics multiple steps in prostate cancer progression, was also derived from RWPE-1 cells by exposure to N-methyl-N-nitrosourea (MNU). See the WPE1-NA22 (ATCC CRL-2849), WPE1-NB14 (ATCC CRL-2850, WPE1-NB11 (ATCC CRL-2851) and WPE1-NB26 (ATCC CRL-2852) cell lines.

The depositor reports that the RWPE-1 cell line (TS210976) was screened, and found negative for, Hepatitis B virus, Hepatitis C virus and Human immunodeficiency virus.

ATCC confirmed this cell line is positive for the presence of HPV viral DNA sequences via PCR.

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbeccos phosphate-buffered saline (D-PBS).
3. Add 2.0 to 3.0 mL (to a T-25 flask) or 3.0 to 4.0 mL (to a T-75 flask) of 0.05% Trypsin - 0.53mM EDTA solution, diluted 1:1 with D-PBS, and place flask in a 37°C incubator for 5 to 8 minutes. Observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 10 minutes).
   Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach.
4. Add 6.0 to 8.0 mL of 0.1% Soybean Trypsin Inhibitor (ATCC^® 30-2014™) or 2% fetal bovine serum in D-PBS, as appropriate, and aspirate cells by gently pipetting.
5. Transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 7 minutes.
6. Discard supernatant and resuspend cells in fresh serum-free growth medium. Add appropriate aliquots of cell suspension to new culture vessels. An inoculum of 2 X 10⁴ to 4 X 10⁴ viable cells/cm² is recommended.
7. Incubate cultures at 37°C. We recommend that you maintain cultures at a cell concentration between 4 X 10⁴ and 7 X 10^4xa0cells/cm².

Webber MM, Rhim JS. Immortalized and malignant human prostatic cell lines. US Patent 5,824,488 dated Oct 20 1998

Bello D, et al. Androgen responsive adult human prostatic epithelial cell lines immortalized by human papillomavirus 18. Carcinogenesis 18: 1215-1223, 1997. PubMed: 9214605

Webber MM, et al. Acinar differentiation by non-malignant immortalized human prostatic epithelial cells and its loss by malignant cells. Carcinogenesis 18: 1225-1231, 1997. PubMed: 9214606

Okamoto M, et al. Interleukin-6 and epidermal growth factor promote anchorage-independent growth of immortalized human prostatic epithelial cells treated with N-methyl-N-nitrosourea. Prostate 35: 255-262, 1998. PubMed: 9609548

Webber MM, et al. Immortalized and tumorigenic adult human prostatic epithelial cell lines: characteristics and applications. Part I. Cell markers and immortalized nontumorigenic cell lines. Prostate 29: 386-394, 1996. PubMed: 8977636

Webber MM, et al. Immortalized and tumorigenic adult human prostatic epithelial cell lines: characteristics and applications Part 2. Tumorigenic cell lines. Prostate 30: 58-64, 1997. PubMed: 9018337

Webber MM, et al. Immortalized and tumorigenic adult human prostatic epithelial cell lines: characteristics and applications. Part 3. Oncogenes, suppressor genes, and applications. Prostate 30: 136-142, 1997. PubMed: 9051152

Kremer R, et al. ras Activation of human prostate epithelial cells induces overexpression of parathyroid hormone-related peptide. Clin. Cancer Res. 3: 855-859, 1997. PubMed: 9815759

Jacob K, et al. Osteonectin promotes prostate cancer cell migration and invasion: a possible mechanism for metastasis to bone. Cancer Res. 59: 4453-4457, 1999. PubMed: 10485497

Achanzar WE, et al. Cadmium induces c-myc, p53, and c-jun expression in normal human prostate epithelial cells as a prelude to apoptosis. Toxicol. Appl. Pharmacol. 164: 291-300, 2000. PubMed: 10799339

Achanzar WE, et al. Cadmium-induced malignant transformation of human prostate epithelial cells. Cancer Res. 61: 455-458, 2001. PubMed: 11212230

Bello-DeOcampo D, et al. Laminin-1 and alpha6beta1 integrin regulate acinar morphogenesis of normal and malignant human prostate epithelial cells. Prostate 46: 142-153, 2001. PubMed: 11170142

Webber MM, et al. Human cell lines as an in vitro/in vivo model for prostate carcinogenesis and progression. Prostate 47: 1-13, 2001. PubMed: 11304724

Epithelial cells from a histologically normal adult human prostate were isolated and subsequently transfected with a plasmid carrying one copy of the human papillomavirus 18 (HPV-18) genome to establish the RWPE-1 (TS210976) cell line.

upon exposure to androgen

Quader ST, et al. Evaluation of the chemopreventive potential of retinoids using a novel in vitro human prostate carcinogenesis model. Mutat. Res. 496: 153-161, 2001. PubMed: 11551491

upregulated upon exposure to androgen

Bello D, et al. Androgen responsive adult human prostatic epithelial cell lines immortalized by human papillomavirus 18. Carcinogenesis 18: 1215-1223, 1997. PubMed: 9214605