Tumor Biology

, 32:1097 | Cite as

Androgen receptor W741C and T877A mutations in AIDL cells, an androgen-independent subline of prostate cancer LNCaP cells

  • Takashi Otsuka
  • Kazuhiro Iguchi
  • Kazuhiro Fukami
  • Kenichiro Ishii
  • Shigeyuki Usui
  • Yoshiki Sugimura
  • Kazuyuki Hirano
Research Article


The androgen-independent LNCaP (AIDL) cell line was generated by maintaining prostate cancer LNCaP cells in a hormone-deprived medium. Notably, synthetic androgen R1881-related gene response is attenuated in AIDL cells as compared to the parental LNCaP cells. The aim of this study was to clarify the mechanisms underlying androgen sensitivity in AIDL cells. We first examined the expression of androgen receptor (AR) and its co-regulators. However, no significant difference in mRNA expression was found between LNCaP and AIDL cells. Remarkably, AR protein levels were induced by R1881 and DHT in LNCaP cells, but not in AIDL cells. We next performed the cDNA sequencing to detect mutations in the AR gene. The T877A mutation was detected both in LNCaP and AIDL cells. Furthermore, AIDL cells harbored a missense substitution (TGG → TGT) in the AR gene, which caused a point mutation at codon 741 (W741C). Double T877A and W741C AR mutants have been previously reported to exhibit reduced androgen sensitivity. Hence, the low-androgen-sensitive responses of AIDL cells may be explained, at least in part, by AR gene mutations.


LNCaP AIDL Androgen sensitivity Androgen receptor Mutation 



This study was supported by a Grant-in-Aid from the Uehara Memorial Foundation and a Grant-in-Aid for Young Scientists (No. 21791514) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.


  1. 1.
    Huggins C, Hodges CV. Studies on prostatic cancer. I: The effect of castration, of estrogen and of androgen injection on serum phosphatases in metastatic carcinoma of the prostate. Cancer Res. 1941;1:293–7.Google Scholar
  2. 2.
    Isaacs JT. New strategies for the medical treatment of prostate cancer. BJU Int. 2005;96 Suppl 2:35–40.PubMedCrossRefGoogle Scholar
  3. 3.
    Suzuki H, Ueda T, Ichikawa T, Ito H. Androgen receptor involvement in the progression of prostate cancer. Endocr Relat Cancer. 2003;10:209–16.PubMedCrossRefGoogle Scholar
  4. 4.
    Thalmann GN, Anezinis PE, Chang SM, Zhau HE, Kim EE, Hopwood VL, et al. Androgen-independent cancer progression and bone metastasis in the LNCaP model of human prostate cancer. Cancer Res. 1994;54:2577–81.PubMedGoogle Scholar
  5. 5.
    Kokontis JM, Hay N, Liao S. Progression of LNCaP prostate tumor cells during androgen deprivation: hormone-independent growth, repression of proliferation by androgen, and role for p27Kip1 in androgen-induced cell cycle arrest. Mol Endocrinol. 1998;12:941–53.PubMedCrossRefGoogle Scholar
  6. 6.
    Hara T, Miyazaki J, Araki H, Yamaoka M, Kanzaki N, Kusaka M, et al. Novel mutations of androgen receptor: a possible mechanism of bicalutamide withdrawal syndrome. Cancer Res. 2003;63:149–53.PubMedGoogle Scholar
  7. 7.
    Ishikura N, Kawata H, Nishimoto A, Nakamura R, Ishii N, Aoki Y. Establishment and characterization of an androgen receptor-dependent, androgen-independent human prostate cancer cell line, LNCaP-CS10. Prostate. 2010;70:457–66.PubMedGoogle Scholar
  8. 8.
    Iguchi K, Ishii K, Nakano T, Otsuka T, Usui S, Sugimura Y, et al. Isolation and characterization of LNCaP sublines differing in hormone sensitivity. J Androl. 2007;28:670–8.PubMedCrossRefGoogle Scholar
  9. 9.
    Onishi T, Yamakawa K, Franco OE, Kawamura J, Watanabe M, Shiraishi T, et al. Mitogen-activated protein kinase pathway is involved in alpha6 integrin gene expression in androgen-independent prostate cancer cells: role of proximal Sp1 consensus sequence. Biochim Biophys Acta. 2001;1538:218–27.PubMedCrossRefGoogle Scholar
  10. 10.
    Iguchi K, Otsuka T, Usui S, Ishii K, Onishi T, Sugimura Y, et al. Zinc and metallothionein levels and expression of zinc transporters in androgen-independent subline of LNCaP cells. J Androl. 2004;25:154–61.PubMedGoogle Scholar
  11. 11.
    Ishii K, Imamura T, Iguchi K, Arase S, Yoshio Y, Arima K, et al. Evidence that androgen-independent stromal growth factor signals promote androgen-insensitive prostate cancer cell growth in vivo. Endocr Relat Cancer. 2009;16:415–28.PubMedCrossRefGoogle Scholar
  12. 12.
    Tepper CG, Boucher DL, Ryan PE, Ma AH, Xia L, Lee LF, et al. Characterization of a novel androgen receptor mutation in a relapsed CWR22 prostate cancer xenograft and cell line. Cancer Res. 2002;62:6606–14.PubMedGoogle Scholar
  13. 13.
    Rahman M, Miyamoto H, Chang C. Androgen receptor coregulators in prostate cancer: mechanisms and clinical implications. Clin Cancer Res. 2004;10:2208–19.PubMedCrossRefGoogle Scholar
  14. 14.
    Jaworski T. Degradation and beyond: control of androgen receptor activity by the proteasome system. Cell Mol Biol Lett. 2006;11:109–31.PubMedCrossRefGoogle Scholar
  15. 15.
    Tan J, Sharief Y, Hamil KG, Gregory CW, Zang DY, Sar M, et al. Dehydroepiandrosterone activates mutant androgen receptors expressed in the androgen-dependent human prostate cancer xenograft CWR22 and LNCaP cells. Mol Endocrinol. 1997;11:450–9.PubMedCrossRefGoogle Scholar
  16. 16.
    Gaddipati JP, McLeod DG, Heidenberg HB, Sesterhenn IA, Finger MJ, Moul JW, et al. Frequent detection of codon 877 mutation in the androgen receptor gene in advanced prostate cancers. Cancer Res. 1994;54:2861–4.PubMedGoogle Scholar
  17. 17.
    Taplin ME, Rajeshkumar B, Halabi S, Werner CP, Woda BA, Picus J, et al. Androgen receptor mutations in androgen-independent prostate cancer: Cancer and Leukemia Group B Study 9663. J Clin Oncol. 2003;21:2673–8.PubMedCrossRefGoogle Scholar
  18. 18.
    Urushibara M, Ishioka J, Hyochi N, Kihara K, Hara S, Singh P, et al. Effects of steroidal and non-steroidal antiandrogens on wild-type and mutant androgen receptors. Prostate. 2007;67:799–807.PubMedCrossRefGoogle Scholar
  19. 19.
    Gregory CW, He B, Johnson RT, Ford OH, Mohler JL, French FS, et al. A mechanism for androgen receptor-mediated prostate cancer recurrence after androgen deprivation therapy. Cancer Res. 2001;61:4315–9.PubMedGoogle Scholar
  20. 20.
    Sadar MD. Androgen-independent induction of prostate-specific antigen gene expression via cross-talk between the androgen receptor and protein kinase A signal transduction pathways. J Biol Chem. 1999;274:7777–83.PubMedCrossRefGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2011

Authors and Affiliations

  • Takashi Otsuka
    • 1
    • 2
  • Kazuhiro Iguchi
    • 1
  • Kazuhiro Fukami
    • 1
  • Kenichiro Ishii
    • 3
    • 4
  • Shigeyuki Usui
    • 1
  • Yoshiki Sugimura
    • 3
  • Kazuyuki Hirano
    • 1
  1. 1.Laboratory of PharmaceuticsGifu Pharmaceutical UniversityGifuJapan
  2. 2.Ogaki Municipal HospitalOgakiJapan
  3. 3.Department of Nephro-Urologic Surgery and AndrologyMie University Graduate School of MedicineTsuJapan
  4. 4.Mie University Graduate School of Regional Innovation StudiesTsuJapan

Personalised recommendations