Characterization of fibroblast-free CWR-R1ca castration-recurrent prostate cancer cell line.
The previously established CWR-R1 cell line has been used as an in vitro model representing castration-recurrent prostate cancer. Microscopic observation of subconfluent cells demonstrated two distinct cellular morphologies: polygonal closely aggregated epithelial cells surrounded by bipolar fibroblastic cells with long processes. This study sought to establish and characterize a fibroblast-free derivative of the CWR-R1 cell line.



The CWR-R1ca cell line was established from CWR-R1 cells by removing fibroblasts using multiple cycles of short-term trypsinization, cloning, and pooling single-cell colonies. Authentication of fibroblast-free CWR-R1ca cells was demonstrated by analyzing the expression of cytodifferentiation and prostate-associated markers, DNA and cytogenetic profiling, and growth pattern in the absence or presence of androgen.



CWR-R1ca is an androgen-sensitive cell line that expresses the androgen receptor (AR) and its splice variant 7 and the luminal epithelia markers, CK-8, CK-18, and c-Met. CWR-R1fb fibroblasts isolated from CWR-R1 cells express AR, hepatocyte growth factor-α, and mouse β-actin but not AR-V7 or epithelial markers. Cytogenetic analysis of CWR-R1ca cells revealed a hyperdiploid male with numerical gains in chromosomes 1, 7, 8, 10, 11, and 12, deletion of one chromosome 2 allele, structural abnormalities that include der(1)t(1:4), der(4)t(2:4), der(10)t(4:10), and an unbalanced reciprocal translocation between chromosome 6 and 14. DNA-profiling revealed that CWR-R1ca cells had significant short-tandem repeat marker homology with CWR22Pc and CWR22Rv1 cell lines, which indicated lineage derivation from CWR22 prostate cancer xenografts. CWR-R1ca cells were responsive to the growth stimulatory effects of dihydrotestosterone (DHT) in the femtomolar range.



This study establishes CWR-R1ca cells as a fibroblast-free derivative of the castration-recurrent CWR-R1 cell line. Prostate 76:1067-1077, 2016. © 2016 Wiley Periodicals, Inc.
Department of Cancer Genetics, Center for Genetics and Pharmacology, Roswell Park Cancer Institute, Buffalo, New York.
Mojgan
M
Shourideh
Department of Cancer Genetics, Center for Genetics and Pharmacology, Roswell Park Cancer Institute, Buffalo, New York.
Adam
A
DePriest
Department of Urology, Roswell Park Cancer Institute, Buffalo, New York.|Department of Urology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.|Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.
James L
JL
Mohler
Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.|Department of Pediatrics, Laboratories for Reproductive Biology, University of North Carolina, Chapel Hill, North Carolina.|Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, North Carolina.
Elizabeth M
EM
Wilson
Department of Cancer Genetics, Center for Genetics and Pharmacology, Roswell Park Cancer Institute, Buffalo, New York.|Department of Urology, Roswell Park Cancer Institute, Buffalo, New York.
Shahriar
S
Koochekpour
D000199:Actins / Q000032:analysis; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000379:methods; D002869:Chromosome Aberrations; D002872:Chromosome Deletion; D016172:DNA Fingerprinting; D005347:Fibroblasts / Q000166:cytology*; D015870:Gene Expression; D017228:Hepatocyte Growth Factor / Q000032:analysis; D064593:Heterografts; D006801:Humans; D007621:Karyotyping; D008297:Male; D051379:Mice; D041681:NIH 3T3 Cells; D009368:Neoplasm Transplantation; D011467:Prostate / Q000737:chemistry; D011471:Prostatic Neoplasms / Q000235:genetics / Q000473:pathology; D064129:Prostatic Neoplasms, Castration-Resistant / Q000235:genetics / Q000473:pathology*; D020033:Protein Isoforms / Q000032:analysis; D011944:Receptors, Androgen / Q000032:analysis / Q000235:genetics
D000199:Actins; D015415:Biomarkers; D020033:Protein Isoforms; D011944:Receptors, Androgen; D017228:Hepatocyte Growth Factor
CWR-R1; androgen receptor; prostate cancer