Loss of androgen receptor-dependent growth suppression by prostate cancer cells can occur independently from acquiring oncogenic addiction to androgen receptor signaling
Authors: Donald J Vander Griend
Lizamma Antony
Jason M D&#x27;Antonio
Susan L Dalrymple
John T Isaacs
George Ndikuyeze
Shahriar Koochekpour
Journal: PLoS ONE (2010)
Discipline: Multidisciplinary
Keywords: androgen receptor signaling; prostate cancer cells; AR mutation; AR amplification; AR over-expression
Affiliations: Department of Oncology, The Sidney Kimmel Comprehensive Cancer Center, The Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America. 

Abstract:  The conversion of androgen receptor (AR) signaling as a mechanism of growth suppression of normal prostate epithelial cells to that of growth stimulation in prostate cancer cells is often associated with AR mutation, amplification and over-expression. Thus, down-regulation of AR signaling is commonly therapeutic for prostate cancer. The E006AA cell line was established from a hormone naïve, localized prostate cancer. E006AA cells are genetically aneuploid and grow equally well when xenografted into either intact or castrated male NOG but not nude mice. These cells exhibit: 1) X chromosome duplication and AR gene amplification, although paradoxically not coupled with increased AR expression, and 2) somatic, dominant-negative Serine-599-Glycine loss-of-function mutation within the dimerization surface of the DNA binding domain of the AR gene. No effect on the growth of E006AA cells is observed using targeted knockdown of endogenous mutant AR, ectopic expression of wild-type AR, or treatment with androgens or anti-androgens. E006AA cells represent a prototype for a newly identified subtype of prostate cancer cells that exhibit a dominant-negative AR loss-of-function in a hormonally naïve patient. Such loss-of-function eliminates AR-mediated growth suppression normally induced by normal physiological levels of androgens, thus producing a selective growth advantage for these malignant cells in hormonally naïve patients. These data highlight that loss of AR-mediated growth suppression is an independent process, and that, without additional changes, is insufficient for acquiring oncogene addiction to AR signaling. Thus, patients with prostate cancer cells harboring such AR loss-of-function mutations will not benefit from aggressive hormone or anti-AR therapies even though they express AR protein. 

Retracted: No

PubPeer comments:
 Prostate cancer in African Americans is much more lethal than in Europeans or European Americans. As all prostate cancer cell lines available had been isolated from European American patients, research into African American prostate cancer was significantly impeded, until the E006AA cell line was reported as the first prostate cancer cell line of African American origin. This was celebrated as a major development for cancer research. Unfortunately, it was later found that this cell line is neither of African American nor of prostate origin. It is a misidentified cell line, most likely a subclone of the 786-O renal adenocarcinoma cell line. The STR profile of the E006AA subclone E006AA-hT is available at the American Type Culture Collection (ATCC) site: https: www.atcc.org en Products Cells_and_Microorganisms By_Tissue Prostate CRL-3277.aspx#specifications. All these STRs match the profile of 786-O cells (also available at the ATCC site): https: www.atcc.org Products All CRL-1932.aspx#specifications (even though 2 STRs were lost, probably during the selection of the E006AA-hT subclone from the E006AA cells, the statistical probability based on these STR profiles is that the E006AA and 786-O cells originated from the same patient. Considering that the 786-O cell line was isolated in the 1970s and the E006AA cells about 30 years later, it is unlikely that the same patient was seen in 2 different hospitals with 2 different cancers 30 years apart). E006AA cells are not of African American origin either. Ancestry testing showed that the cells are from a European American patient: See Table 1 in https: www.semanticscholar.org paper RNA-Seq-Reveals-Genetic-Differences-in-African-and-Bernot-Cauley cb63591b77a4cb8dae4533719e552a316402b5ae. Not surprisingly, the 786-O renal adenocarcinoma cell line is well known to have been isolated from a European American patient. How could this mix-up have happened? Were the 786-O cells present in the same Urology lab (at Louisiana State University) where the E006AA cells were first reported https: www.ncbi.nlm.nih.gov pubmed 15162380? Is there any evidence of that? Yes: https: www.ncbi.nlm.nih.gov pubmed 12131366. So for the past 15 years, the E006AA cells have been used extensively as an u201cAfrican American prostate cancer cell lineu201d, filling the literature with confusing and irrelevant results. For example, D Antonio JM, et al PLoS One. 2010 Jul 8;5(7):e11475, found that no effect on the growth of E006AA cells is observed using targeted knockdown of endogenous androgen receptor, ectopic expression of wild-type androgen receptor, or treatment with androgens or anti-androgens. This would be an unexpected result for a prostate cancer cell line, but not surprising for a renal adenocarcinoma cell line. ATCC, despite its rhetoric on cell authentication, continues to sell the E006AA-hT cells as u201cAfrican American prostate canceru201d, resulting in many more publications that are simply wrong. Isnu2019t it time the ATCC stopped selling a renal adenocarcinoma cell line as a model of u201cAfrican American prostate canceru201d to unsuspecting researchers? How can we find a cure for prostate cancer by studying renal adenocarcinoma cells? https: www.atcc.org Products All CRL-3277.aspx , 
 https: www.ncbi.nlm.nih.gov pubmed 30787054 More proof that the E006AA cells are neither an African American nor a prostate cancer cell line. They are a kidney cancer cell line from a white male. And yet, ATCC continues to sell them as a prostate cancer model and cites the papers that erroneously claim it is of African ancestry, misguiding researchers and causing great confusion in the literature. , 
 https: onlinelibrary.wiley.com doi full 10.1002 pros.23800  Correction: Establishment and characterization of a primary androgenu2010responsive Africanu2010American prostate cancer cell line, E006AA. Prostate 2004;60(2):145u2010152 New information has been obtained on the E006AA epithelial cell line which was characterized in the paper we published in the Prostate 60(2):141u2010152 (2004) as having originated from an African American prostate tumor line. We now believe that this cell line is a subclone of the 786u2010O renal adenocarcinoma line from a white male. The STR profile of the E006AA subclone E006AAu2010hT, available at the American Type Culture Collection (ATCC) site has STRs which match the profile of 786u2010O cells (also available at the ATCC site). The analyses presented in the 2004 article are original and accurate using the techniques and methodologies that were available at the time (eg, H E staining, cellu2010type and prostateu2010specific marker, AR expression, RT_PCR, Tumorigenicity, Karyotype, etc). We had no reason to doubt that the line we cultured did not originate from an African American prostate tumor. We now believe it may have been contaminated in its early passages with the 786u20100 cell line before it was cryopreserved, as 786u20100 cells were also studied in the lab from which this line originated. We urge ATCC to cease advertising this cell line, E006AAu2010hT, as originating from African American prostate cancer tissue. We would also like to apologize for the confusion, costs, and delays in research this mistake may have caused other researchers; and we thank the researchers who brought this error to our attention. ,