Like other malignancies, prostate tumors are thought to contain cancer stem-like cells (CSCs) that are responsible for growth, metastasis, and therapy resistance. ΔNp63 (also called p40) is a regulator of normal prostate stem/progenitor cell activities and a marker of normal basal epithelial cells. The levels of ΔNp63 are reduced in prostate adenocarcinomas, although there is also evidence that ΔNp63 is involved in CSC regulation and drives metastasis to the bone. We studied metastatic deposits of prostate cancers with isoform-specific ΔNp63 and TAp63 antibodies. We identified p63-positive cells in only 3 of 42 metastatic prostate tumors (7%), including 2/38 (5.3%) “usual-type” adenocarcinomas. ΔNp63 and TAp63 isoforms were present in the nuclei of a small subpopulation (< 1%) of tumor cells in these metastases. ΔNp63-positive cells showed a basal-like cell phenotype (cytokeratin 8- and androgen receptor-negative, high molecular weight cytokeratin- and cytokeratin 19-positive), distinct from the tumor bulk. TAp63-positive cells were similar but were sometimes cytokeratin 8-positive. A subset of ΔNp63-positive tumor cells were CD44-positive, a marker of “basal” CSCs but were not positive for the “epithelial” CSC marker ALDH1. TAp63 was not associated with either of these CSC markers. None of the tumors containing p63-positive cells showed evidence of bone metastasis, compared with 28% of the p63-negative tumors. These data show that both ΔNp63 and TAp63 are present in only a small proportion of prostate adenocarcinomas and do not associate with metastasis. The data suggest heterogeneity of CSCs in prostate cancer, similar to other cancer types.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Scanned digital slide images from which the photomicrographs in this article are derived are available on request.
Shen MM, Rubin MA (2019) Prostate cancer research at the crossroads. Cold Spring Harb Perspect Med 9:a036277. https://doi.org/10.1101/cshperspect.a036277
Wang G, Zhao D, Spring DJ, DePinho RA (2018) Genetics and biology of prostate cancer. Genes Dev 32:1105–1140. https://doi.org/10.1101/gad.315739.118
Li JJ, Shen MM (2019) Prostate stem cells and cancer stem cells. Cold Spring Harb Perspect Med 9:a030395. https://doi.org/10.1101/cshperspect.a030395
Ali HR, Dawson S-J, Blows FM, Provenzano E, Pharoah PD, Caldas C (2011) Cancer stem cell markers in breast cancer: pathological, clinical and prognostic significance. Breast Cancer Res 13:R118. https://doi.org/10.1186/bcr3061
Liu Y, Nenutil R, Appleyard MV, Boylan M, Thompson AM, Coates PJ (2014) Lack of correlation of stem cell markers in breast cancer stem cells. Br J Cancer 110:2063–2071. https://doi.org/10.1038/bjc.2014.105
Hsu C-L, Chung F-H, Chen C-H, Hsu TT, Liu SM, Chung DS, Hsu YF, Chen CL, Ma N, Lee HC (2016) Genotypes of cancer stem cells characterized by epithelial-to-mesenchymal transition and proliferation related functions. Sci Rep 6:32523. https://doi.org/10.1038/srep32523
Sampayo RG, Bissell MJ (2019) Cancer stem cells in breast and prostate: fact or fiction? Adv Cancer Res 144:315–341. https://doi.org/10.1016/bs.acr.2019.03.010
Crum CP, McKeon FD (2010) p63 in epithelial survival, germ cell surveillance, and neoplasia. Annu Rev Pathol 5:349–371. https://doi.org/10.1146/annurev-pathol-121808-102117
Nekulova M, Holcakova J, Coates P, Vojtesek B (2011) The role of p63 in cancer, stem cells and cancer stem cells. Cell Mol Biol Lett 16:296–327. https://doi.org/10.2478/s11658-011-0009-9
Orzol P, Holcakova J, Nekulova M, Nenutil R, Vojtesek B, Coates PJ (2015) The diverse oncogenic and tumour suppressor roles of p63 and p73 in cancer: a review by cancer site. Histol Histopathol 30:503–521. https://doi.org/10.14670/HH-30.503
Melino G, Memmi EM, Pelicci PG, Bernassola F (2015) Maintaining epithelial stemness with p63. Sci Sig 8:re9. https://doi.org/10.1126/scisignal.aaa1033
Signoretti S, Waltregny D, Dilks J, Isaac B, Lin D, Garraway L, Yang A, Montironi R, McKeon F, Loda M (2000) p63 is a prostate basal cell marker and is required for prostate development. Am J Pathol 157:1769–1775. https://doi.org/10.1016/S0002-9440(10)64814-6
Signoretti S, Pires MM, Lindauer M, Horner JW, Grisanzio C, Dhar S, Majumder P, McKeon F, Kantoff PW, Sellers WR, Loda M (2005) p63 regulates commitment to the prostate cell lineage. Proc Natl Acad Sci U S A 102:11355–11360. https://doi.org/10.1073/pnas.0500165102
Pignon J-C, Grisanzio C, Geng Y, Song J, Shivdasani R, Signoretti S (2013) p63-expressing cells are the stem cells of developing prostate, bladder, and colorectal epithelia. Proc Natl Acad Sci U S A 110:8105–8110. https://doi.org/10.1073/pnas.1221216110
Huang Y, Hamana T, Liu J, Wang C, An L, You P, Chang JYF, Xu J, McKeehan WL, Wang F (2015) Prostate sphere-forming stem cells are derived from the P63-expressing basal compartment. J Biol Chem 290:17745–17752. https://doi.org/10.1074/jbc.M115.661033
Garraway IP, Sun W, Tran CP, Perner S, Zhang B, Goldstein AS, Hahm SA, Haider M, Head CS, Reiter RE, Rubin MA, Witte ON (2010) Human prostate sphere-forming cells represent a subset of basal epithelial cells capable of glandular regeneration in vivo. Prostate 70:491–501. https://doi.org/10.1002/pros.21083
Portillo-Lara R, Alvarez MM (2015) Enrichment of the cancer stem phenotype in sphere cultures of prostate cancer cell lines occurs through activation of developmental pathways mediated by the transcriptional regulator ΔNp63α. PLoS One 10:e0130118. https://doi.org/10.1371/journal.pone.0130118
Nylander K, Vojtesek B, Nenutil R, Lindgren B, Roos G, Zhanxiang W, Sjöström B, Dahlqvist Å, Coates PJ (2002) Differential expression of p63 isoforms in normal tissues and neoplastic cells. J Pathol 198:417–427. https://doi.org/10.1002/path.1231
Sailer V, Stephan C, Wernert N, Perner S, Jung K, Kristiansen G (2013) Comparison of p40 (ΔNp63) and p63 expression in prostate tissues--which one is the superior diagnostic marker for basal cells? Histopathology 63:50–56. https://doi.org/10.1111/his.12116
Shah RB, Kunju LP, Shen R, LeBlanc M, Zhou M, Rubin MA (2004) Usefulness of basal cell cocktail (34betaE12 + p63) in the diagnosis of atypical prostate glandular proliferations. Am J Clin Pathol 122:517–523. https://doi.org/10.1309/WRM5-1C70-P1NB-FE4K
Ali TZ, Epstein JI (2008) False positive labeling of prostate cancer with high molecular weight cytokeratin: p63 a more specific immunomarker for basal cells. Am J Surg Pathol 32:1890–1895. https://doi.org/10.1097/PAS.0b013e31817ce994
Osunkoya AO, Hansel DE, Sun X, Neto GJ, Epstein JI (2008) Aberrant diffuse expression of p63 in adenocarcinoma of the prostate on needle biopsy and radical prostatectomy: report of 21 cases. Am J Surg Pathol 32:461–467. https://doi.org/10.1097/PAS.0b013e318157020e
Wu A, Kunju LP (2013) Prostate cancer with aberrant diffuse p63 expression: report of a case and review of the literature and morphologic mimics. Arch Pathol Lab Med 137:1179–1184. https://doi.org/10.5858/arpa.2013-0254-CR
Tan H-L, Haffner MC, Esopi DM, Vaghasia AM, Giannico GA, Ross HM, Ghosh S, Hicks JL, Zheng Q, Sangoi AR, Yegnasubramanian S, Osunkoya AO, de Marzo AM, Epstein JI, Lotan TL (2015) Prostate adenocarcinomas aberrantly expressing p63 are molecularly distinct from usual-type prostatic adenocarcinomas. Mod Pathol 28:446–456. https://doi.org/10.1038/modpathol.2014.115
Uchida K, Ross H, Lotan T, Pignon J-C, Signoretti S, Epstein JI, Illei PB (2015) ΔNp63 (p40) expression in prostatic adenocarcinoma with diffuse p63 positivity. Hum Pathol 46:384–389. https://doi.org/10.1016/j.humpath.2014.11.011
Torres A, Alshalalfa M, Davicioni E, Gupta A, Yegnasubramanian S, Wheelan SJ, Epstein JI, de Marzo AM, Lotan TL (2018) ETS2 is a prostate basal cell marker and is highly expressed in prostate cancers aberrantly expressing p63. Prostate 78:896–904. https://doi.org/10.1002/pros.23646
Rosenbluth JM, Johnson K, Tang L, Triplett, Pientenpol JA (2009) Evaluation of p63 and p73 antibodies for cross-reactivity. Cell Cycle 8:3702–3706. https://doi.org/10.4161/cc.8.22.10036
Nekulova M, Holcakova J, Nenutil R, Stratmann R, Bouchalova P, Müller P, Mouková L, Coates PJ, Vojtesek B (2013) Characterization of specific p63 and p63-N-terminal isoform antibodies and their application for immunohistochemistry. Virchows Arch 463:415–425. https://doi.org/10.1007/s00428-013-1459-4
Lee AHS (2013) Use of immunohistochemistry in the diagnosis of problematic breast lesions. J Clin Pathol 66:471–477. https://doi.org/10.1136/jclinpath-2012-201109
Ribeiro-Silva A, Ramalho LNZ, Garcia SB, Brandao DF, Chahud F, Zucoloto S (2005) p63 correlates with both BRCA1 and cytokeratin 5 in invasive breast carcinomas: further evidence for the pathogenesis of the basal phenotype of breast cancer. Histopathology 47:458–466. https://doi.org/10.1111/j.1365-2559.2005.02249.x
Liu Y, Nekulova M, Nenutil R, Horakova I, Appleyard MV, Murray K, Holcakova J, Galoczova M, Quinlan P, Jordan LB, Purdie CA, Vojtesek B, Thompson AM, Coates PJ (2020) ∆Np63/p40 correlates with the location and phenotype of basal/mesenchymal cancer stem-like cells in human ER+ and HER2+ breast cancers. J Pathol Clin Res 6:83–93. https://doi.org/10.1002/cjp2.149
Kim J, Villadsen R, Sørlie T et al (2012) Tumor initiating but differentiated luminal-like breast cancer cells are highly invasive in the absence of basal-like activity. Proc Natl Acad Sci U S A 109:6124–6129. https://doi.org/10.1073/pnas.1203203109
Maitland NJ, Collins AT (2008) Prostate cancer stem cells: a new target for therapy. J Clin Oncol 26:2862–2870. https://doi.org/10.1200/JCO.2007.15.1472
Parsons JK, Gage WR, Nelson WG, De Marzo AM (2001) p63 protein expression is rare in prostate adenocarcinoma: implications for cancer diagnosis and carcinogenesis. Urology 58:619–624. https://doi.org/10.1016/s0090-4295(01)01311-5
Resetkova E, Reis-Filho JS, Jain RK, Mehta R, Thorat MA, Nakshatri H, Badve S (2010) Prognostic impact of ALDH1 in breast cancer: a story of stem cells and tumor microenvironment. Breast Cancer Res Treat 123:97–108. https://doi.org/10.1007/s10549-009-0619-3
Epstein JI, Egevad L, Humphrey PA, Montroni R (2014) Best practices recommendations in the application of immunohistochemistry in the prostate: report from the International Society of Urologic Pathology consensus conference. Am J Surg Pathol 38:e6–e19. https://doi.org/10.1097/PAS.0000000000000238
Reiner T, de Las PA, Parrondo R, Perez-Stable C (2007) Progression of prostate cancer from a subset of p63-positive basal epithelial cells in FG/Tag transgenic mice. Mol Cancer Res 5:1171–1179. https://doi.org/10.1158/1541-7786.MCR-07-0024
Di Giacomo V, Tian TV, Mas A et al (2017) ΔNp63α promotes adhesion of metastatic prostate cancer cells to the bone through regulation of CD82. Oncogene 36:4381–4392. https://doi.org/10.1038/onc.2017.42
Gu G, Yuan J, Wills M, Kasper S (2007) Prostate cancer cells with stem cell characteristics reconstitute the original human tumor in vivo. Cancer Res 67:4807–4815. https://doi.org/10.1158/0008-5472.CAN-06-4608
Kurita T, Medina RT, Mills AA, Cunha GR (2004) Role of p63 and basal cells in the prostate. Development 131:4955–4964. https://doi.org/10.1242/dev.01384
Tucci P, Agostini M, Grespi F, Markert EK, Terrinoni A, Vousden KH, Muller PAJ, Dötsch V, Kehrloesser S, Sayan BS, Giaccone G, Lowe SW, Takahashi N, Vandenabeele P, Knight RA, Levine AJ, Melino G (2012) Loss of p63 and its microRNA-205 target results in enhanced cell migration and metastasis in prostate cancer. Proc Natl Acad Sci U S A 109:15312–15317. https://doi.org/10.1073/pnas.1110977109
Hudson DL, Guy AT, Fry P, O'Hare MJ, Watt FM, Masters JR (2001) Epithelial cell differentiation pathways in the human prostate: identification of intermediate phenotypes by keratin expression. J Histochem Cytochem 49:271–278. https://doi.org/10.1177/002215540104900214
Wang Y, Hayward S, Cao M, Thayer K, Cunha G (2001) Cell differentiation lineage in the prostate. Differentiation 68:270–279
Liu S, Cong Y, Wang D, Sun Y, Deng L, Liu Y, Martin-Trevino R, Shang L, McDermott SP, Landis MD, Hong S, Adams A, D’Angelo R, Ginestier C, Charafe-Jauffret E, Clouthier SG, Birnbaum D, Wong ST, Zhan M, Chang JC, Wicha MS (2014) Breast cancer stem cells transition between epithelial and mesenchymal states reflective of their normal counterparts. Stem Cell Reports 2:78–91. https://doi.org/10.1016/j.stemcr.2013.11.009
Coates PJ, Nenutil R, Holcakova J, Nekulova M, Podhorec J, Svoboda M, Vojtesek B (2018) p63 isoforms in triple-negative breast cancer: ΔNp63 associates with the basal phenotype whereas TAp63 associates with androgen receptor, lack of BRCA mutation, PTEN and improved survival. Virchows Arch 472:351–359. https://doi.org/10.1007/s00428-018-2324-2
This work was supported by the Czech Science Foundation (19-06530S), the European Regional Development Fund - Project ENOCH (No. CZ.02.1.01/0.0/0.0/16_019/0000868) and the Ministry of Health, Czech Republic—conceptual development of research organization (MMCI, 00209805). The MMCI biobank is supported by grant LM2018125 from the Ministry of Education, Youth and Sports and co-funded by ADOPT BBMRI-ERIC, supported by EU Horizon 2020 (grant agreement No. 676550). MG was a Brno Ph.D. Talent Scholarship Holder, funded by the Brno City Municipality.
Conflict of interest
BV is a consultant for and RN is a co-owner of Moravian Biotechnology, who originally produced the p63 monoclonal antibodies used in this study. The company did not provide financial support or have any influence over the design, execution, or interpretation of the data. MG, ZP, and PJC declare that they have no competing interests.
The study was performed retrospectively on redundant excess tissues. All patient data were anonymized. In accordance with the Declaration of Helsinki and the regulations of the European Union, the use of excess and redundant tissues was approved following local ethical committee review by the Biobanking and Biomolecular resources Research Infrastructure (BBMRI) at the Masaryk Memorial Cancer Institute, Brno. Patient consent is not required for the retrospective use of anonymized, redundant excess tissues.
Consent to participate
Consent for publication
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Galoczova, M., Nenutil, R., Pokorna, Z. et al. TAp63 and ΔNp63 (p40) in prostate adenocarcinomas: ΔNp63 associates with a basal-like cancer stem cell population but not with metastasis. Virchows Arch (2020). https://doi.org/10.1007/s00428-020-02944-z
- Prostate cancer
- Cancer stem cells