Annals of Surgical Oncology

, Volume 5, Issue 4, pp 342–349 | Cite as

Cell cycle regulation of human pancreatic cancer by tamoxifen

  • Emily K. Robinson
  • Ana M. Grau
  • Douglas B. Evans
  • Christine M. Smid
  • Paul J. Chiao
  • James L. Abbruzzese
  • Elizabeth A. Grimm
Original Articles


Background: Clinical trials have suggested a survival advantage for selected patients with metastatic pancreatic cancer treated with tamoxifen. We sought to identify the molecular mechanism by which tamoxifen inhibits human pancreatic cancer cell (HPCC) growth.

Methods: HPCCs were grown in tamoxifen and growth inhibition was determined by3H-thymidine uptake and by the MTT assay; changes in cell viability were determined by cell counts. Cell cycle alterations were evaluated by FACS, and the induction of apoptosis was evaluated using the TUNEL assay. Total cellular RNA was isolated after tamoxifen treatment, and Northern blot analysis was performed for p21 waf1 .

Results: Tamoxifen inhibited HPCC growth as measured by inhibition of3H-thymidine incorporation and by the MTT assay. However, there was no decrease in the total number of viable cells after 6 days of treatment with 10 µM of tamoxifen and no evident apoptosis, confirming the absence of a cytotoxic effect. Cell cycle analysis revealed cellular arrest in the G0/G1 phase, which correlated with p21 waf1 mRNA upregulation in response to tamoxifen treatment.

Conclusions: Tamoxifen inhibits HPCC growth by inducing G0/G1 arrest with an associated increase in p21 waf1 mRNA expression. Tamoxifen is an effective inhibitor of HPCC growth in vitro and warrants further in vivo study.

Key Words

Cell lines Apoptosis Cell cycle arrest p21waf1 


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  1. 1.
    Pousette A, Carlstrom K, Skoldefors H, et al. Purification and partial characterization of a 17β-estradiol binding macromolecule in the human pancreas.Cancer Res 1982;42:633–7.PubMedGoogle Scholar
  2. 2.
    Benez C, Hollander C, Miller B. Endocrine-responsive pancreatic carcinoma: steroid binding and cytotoxicity in human tumor cell lines.Cancer Res 1986;46:2276–81.Google Scholar
  3. 3.
    Theve NO, Pousette A, Carlstrom K. Adenocarcinoma of the pancreas—a hormone sensitive tumor? A preliminary report of Nolvadex treatment.Clin Oncol 1983;9:193–7.PubMedGoogle Scholar
  4. 4.
    Tonnesen K, Kamp-Jensen M. Antiestrogen therapy in pancreatic carcinoma: a preliminary report.Eur J Surg Oncol 1986;12:69–70.PubMedGoogle Scholar
  5. 5.
    Wong A, Chan A. Survival benefit of tamoxifen therapy in adenocarcinoma of the pancreas. A case-control study.Cancer 1993;71:2200–3.PubMedGoogle Scholar
  6. 6.
    Taylor OM, Benson EA, McMahon MJ. Clinical trial of tamoxifen in patients with irresectable pancreatic adenocarcinoma.Br J Surg 1993;80:384–6.PubMedGoogle Scholar
  7. 7.
    Keating JJ, Johnson PJ, Cochrane AMG, et al. A prospective randomised controlled trial of tamoxifen and cyproterone acetate in pancreatic carcinoma.Br J Cancer 1989;60:789–92.PubMedGoogle Scholar
  8. 8.
    Bakkevold KE, Pettersen A, Arnesjo B, Espehaug B. Tamoxifen therapy in unresectable adenocarcinoma of the pancreas and the papilla of Vater.Br J Surg 1990;77:25–30.Google Scholar
  9. 9.
    Woods CM, Zhu J, McQueney PA, Bollag D, Lazarides E. Taxol-induced mitotic block triggers rapid onset of a p53-independent apoptotic pathway.Mol Med 1995;1:505–26.Google Scholar
  10. 10.
    Danesi R, Figg WD, Reed E, Myers CE. Paclitaxel (taxol) inhibits protein isoprenylation and induces apoptosis in PC-3 human prostate cancer cells.Mol Pharm 1995;47:1106–11.Google Scholar
  11. 11.
    Jordon MA, Wendell K, Gardiner S, Derry WB, Copp H, Wilson L. Mitotic block induced in HeLa cells by low concentrations of paclitaxel (taxol) results in abnormal mitotic exit and apoptotic cell death.Cancer Res 1996;56:816–25.Google Scholar
  12. 12.
    Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidium thiocyanate-phenol-chloroform extraction.Anal Biochem 1987;162:156–9.CrossRefPubMedGoogle Scholar
  13. 13.
    Sambrook J, Fritsch E, Maniatis T. Synthesis of uniformly labeled DNA probes using random oligonucleotide primers. In: Nolan C, ed.Molecular Cloning. A Laboratory Manual. Cold Spring Harbor, NY: Laboratory Press, 1989;2:10.13–5.Google Scholar
  14. 14.
    Sherr CJ. G1 phase progression: cycling on cue.Cell 1994;79:551–5.CrossRefPubMedGoogle Scholar
  15. 15.
    McClay EF, Jones JA, Winski PJ, Albright KD, Christen RD, Howell SB. Determinants of tamoxifen sensitivity control the nature of the synergistic interaction between tamoxifen and cisplatin.Cancer Res 1996;56:3993–7.PubMedGoogle Scholar
  16. 16.
    Perry RR, Kang Y, Greaves B. Effects of tamoxifen on growth and apoptosis of estrogen-dependent and independent human breast cancer cells.Ann Surg Oncol 1995;2:238–45.PubMedGoogle Scholar
  17. 17.
    Michieli P, Chedid M, Lin D, Pierce JH, Mercer EW, Givol D. Induction ofWAF1/CIP1 by a p53-independent pathway.Cancer Res 1994;54:3391–5.PubMedGoogle Scholar
  18. 18.
    Li X-S, Rishi AK, Shao Z-M, et al. Posttranscriptional regulation of p21WAF1/CIP1 expression in human breast carcinoma cells.Cancer Res 1996;56:5055–62.PubMedGoogle Scholar
  19. 19.
    Ruggeeri B, Zhang S-Y, Caamano J, DiRado M, Flynn SD, Klein-Szanto AJP. Human pancreatic carcinomas and cell lines reveal frequent and multiple alterations in p53 and Rb-1 tumor-suppressor genes.Oncogene 1992;7:1503–11.Google Scholar
  20. 20.
    Pan G, Ying W, Vickers SM, Phillips JO, McDonald JM. Involvement of the Fas/APO-1(CD95) in tamoxifen-induced apoptosis in cholangiocarcinoma.Proceedings of the American Association of Cancer Research 1997;38:625.Google Scholar
  21. 21.
    Reddel RR, Murphy LC, Hall RE, Sutherland RL. Differential sensitivity of human breast cancer cell lines to the growth-inhibitory effects of tamoxifen.Cancer Res 1985;45:1525–31.PubMedGoogle Scholar
  22. 22.
    Greenway B, Iqbal M, Johnson P, Williams R. Oestrogen receptor proteins in malignant and fetal pancreas.Br Med J 1981;283:751–3.Google Scholar
  23. 23.
    Taylor O, Teasdale P, Cowen P, McMahon M, Benson E. Classical oestrogen receptor is not detectable in pancreatic adenocarcinoma.Br J Cancer 1992;66:503–6.PubMedGoogle Scholar
  24. 24.
    Boctor A, Band P, Grossman A. Requirement for an accessory factor for binding [3H]estradiol to protein in the cytosol fraction of rat pancreas.Proc Natl Acad Sci USA 1981;78:5648–51.PubMedGoogle Scholar
  25. 25.
    Kuramoto M, Yamashita J-I, Ogawa M. Tissue-type plasminogen activator predicts endocrine responsiveness of human pancreatic carcinoma cells.Cancer 1995;75:1263–72.PubMedGoogle Scholar
  26. 26.
    McClay E, McClay M-E, Jones J, Winski P, Christen R, Howell S, Hall PA. Phase I study of high dose tamoxifen and weekly cisplatin in patients with metastatic melanoma.Cancer 1997;79:1037–43.CrossRefPubMedGoogle Scholar
  27. 27.
    Stuart N, Philip P, Harris A, et al. High-dose tamoxifen as an effector of etoposide cytotoxicity. Clinical effects and in vitro assessment in p-glycoprotein expressing cell lines.Br J Cancer 1992;66:833–9.PubMedGoogle Scholar
  28. 28.
    Wilking N, Appelgren L-E, Carlstrom K, Pousette A, Theve N. The distribution and metabolism of14C labelled tamoxifen in spayed female mice.Acta Pharmacologica et Toxicologica 1982;50:161–8.PubMedGoogle Scholar

Copyright information

© The Society of Surgical Oncology, Inc. 1998

Authors and Affiliations

  • Emily K. Robinson
    • 3
    • 1
  • Ana M. Grau
    • 1
  • Douglas B. Evans
    • 1
  • Christine M. Smid
    • 3
  • Paul J. Chiao
    • 1
  • James L. Abbruzzese
    • 2
  • Elizabeth A. Grimm
    • 3
    • 1
  1. 1.Department of Surgical OncologyThe University of Texas M. D. Anderson Cancer CenterHouston
  2. 2.Department of Medical OncologyThe University of Texas M. D. Anderson Cancer CenterHouston
  3. 3.Dept. of Tumor BiologyThe University of Texas M. D. Anderson Cancer CenterHouston

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