Tumor Biology

, Volume 37, Issue 9, pp 12133–12140 | Cite as

Expression and clinical significance of Wee1 in colorectal cancer

  • Eivind Valen Egeland
  • Kjersti Flatmark
  • Jahn M. Nesland
  • Vivi Ann Flørenes
  • Gunhild M. Mælandsmo
  • Kjetil BoyeEmail author
Original Article


Wee1 is a nuclear kinase regulating cell cycle progression, and has emerged as a promising therapeutic target in cancer. Expression of Wee1 has been associated with poor outcome in certain tumor types, but the prognostic impact and clinical significance in colorectal cancer is unknown. The expression of Wee1 was examined by immunohistochemistry in primary colorectal carcinomas from a prospectively collected patient cohort, and associations with clinicopathological parameters and outcome were investigated. Cell culture experiments were performed using the cell lines RKO and SW620, and the relationship with the metastasis-promoting protein S100A4 was investigated. Nuclear expression was detected in 229 of the 258 tumors analyzed (89 %). Wee1 staining was associated with low pT stage, but no other significant associations with demographic or histopathological variables were found. Moderate Wee1 staining intensity was a predictor of favorable metastasis-free and overall survival compared to strong intensity and no or weak staining. The fraction of positive cells was not a prognostic factor in the present cohort. Inhibition of Wee1 expression using siRNA or treatment with the Wee1 inhibitor MK-1775 reduced expression of the metastasis-promoting protein S100A4, but no relationship between Wee1 and S100A4 was found in the patient samples. In conclusion, Wee1 is highly expressed in primary colorectal carcinomas, but few relevant associations with clinicopathological parameters or outcome were found. The lack of clinical significance of Wee1 expression could indicate that other tumor types might be better suited for further development of Wee1 inhibitors.


Wee1 S100A4 Colorectal cancer Prognostic marker 



We would like to thank Ellen Hellesylt for excellent technical assistance.

Compliance with ethical standards

The study was approved by the Regional Ethics Committee (no. S-98080) and written informed consent was obtained from the patients.


This work was supported by the Norwegian Cancer Society (grant no. 4218523581 to E.V.E.) and the Research Council of Norway.

Conflicts of interest



  1. 1.
    Do K, Doroshow JH, Kummar S. Wee1 kinase as a target for cancer therapy. Cell Cycle. 2013;12(19):3159–64.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Vriend LE, De Witt Hamer PC, Van Noorden CJ, Wurdinger T. WEE1 inhibition and genomic instability in cancer. Biochim Biophys Acta. 2013;1836(2):227–35.PubMedGoogle Scholar
  3. 3.
    Parker LL, Atherton-Fessler S, Piwnica-Worms H. p107wee1 is a dual-specificity kinase that phosphorylates p34cdc2 on tyrosine 15. Proc Natl Acad Sci U S A. 1992;89(7):2917–21.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Aarts M, Sharpe R, Garcia-Murillas I, Gevensleben H, Hurd MS, Shumway SD, et al. Forced mitotic entry of S-phase cells as a therapeutic strategy induced by inhibition of WEE1. Cancer Discov. 2012;2(6):524–39.CrossRefPubMedGoogle Scholar
  5. 5.
    Dominguez-Kelly R, Martin Y, Koundrioukoff S, Tanenbaum ME, Smits VA, Medema RH, et al. Wee1 controls genomic stability during replication by regulating the Mus81-Eme1 endonuclease. J Cell Biol. 2011;194(4):567–79.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Backert S, Gelos M, Kobalz U, Hanski ML, Bohm C, Mann B, et al. Differential gene expression in colon carcinoma cells and tissues detected with a cDNA array. Int J Cancer. 1999;82(6):868–74.CrossRefPubMedGoogle Scholar
  7. 7.
    Mir SE, De Witt Hamer PC, Krawczyk PM, Balaj L, Claes A, Niers JM, et al. In silico analysis of kinase expression identifies WEE1 as a gatekeeper against mitotic catastrophe in glioblastoma. Cancer Cell. 2010;18(3):244–57.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Magnussen GI, Holm R, Emilsen E, Rosnes AK, Slipicevic A, Florenes VA. High expression of Wee1 is associated with poor disease-free survival in malignant melanoma: potential for targeted therapy. PLoS One. 2012;7(6):e38254.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Slipicevic A, Holth A, Hellesylt E, Trope CG, Davidson B, Florenes VA. Wee1 is a novel independent prognostic marker of poor survival in post-chemotherapy ovarian carcinoma effusions. Gynecol Oncol. 2014;135(1):118–24.CrossRefPubMedGoogle Scholar
  10. 10.
    Yoshida T, Tanaka S, Mogi A, Shitara Y, Kuwano H. The clinical significance of cyclin B1 and Wee1 expression in non-small-cell lung cancer. Ann Oncol. 2004;15(2):252–6.CrossRefPubMedGoogle Scholar
  11. 11.
    De Witt Hamer PC, Mir SE, Noske D, Van Noorden CJ, Wurdinger T. WEE1 kinase targeting combined with DNA-damaging cancer therapy catalyzes mitotic catastrophe. Clin Cancer Res. 2011;17(13):4200–7.CrossRefPubMedGoogle Scholar
  12. 12.
    Mueller S, Haas-Kogan DA. WEE1 kinase as a target for cancer therapy. J Clin Oncol 2015;33(30):3485–7.Google Scholar
  13. 13.
    Carrassa L, Chila R, Lupi M, Ricci F, Celenza C, Mazzoletti M, et al. Combined inhibition of Chk1 and Wee1: in vitro synergistic effect translates to tumor growth inhibition in vivo. Cell Cycle. 2012;11(13):2507–17.CrossRefPubMedGoogle Scholar
  14. 14.
    Davies KD, Cable PL, Garrus JE, Sullivan FX, von Carlowitz I, Huerou YL, et al. Chk1 inhibition and Wee1 inhibition combine synergistically to impede cellular proliferation. Cancer Biol Ther. 2011;12(9):788–96.CrossRefPubMedGoogle Scholar
  15. 15.
    Magnussen GI, Emilsen E, Giller Fleten K, Engesaeter B, Nahse-Kumpf V, Fjaer R, et al. Combined inhibition of the cell cycle related proteins Wee1 and Chk1/2 induces synergistic anti-cancer effect in melanoma. BMC Cancer. 2015;15:462.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Boye K, Maelandsmo GM. S100A4 and metastasis: a small actor playing many roles. Am J Pathol. 2010;176(2):528–35.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Boye K, Nesland JM, Sandstad B, Maelandsmo GM, Flatmark K. Nuclear S100A4 is a novel prognostic marker in colorectal cancer. Eur J Cancer. 2010;46(16):2919–25.CrossRefPubMedGoogle Scholar
  18. 18.
    Egeland EV, Boye K, Pettersen SJ, Haugen MH, Oyjord T, Malerod L et al. Enrichment of nuclear S100A4 during G2/M in colorectal cancer cells: possible association with cyclin B1 and centrosomes. Clin Exp Metastasis 2015;32(8):755–67.Google Scholar
  19. 19.
    Boutros R, Ducommun B. Asymmetric localization of the CDC25B phosphatase to the mother centrosome during interphase. Cell Cycle. 2008;7(3):401–6.CrossRefPubMedGoogle Scholar
  20. 20.
    Jackman M, Lindon C, Nigg EA, Pines J. Active cyclin B1-Cdk1 first appears on centrosomes in prophase. Nat Cell Biol. 2003;5(2):143–8.CrossRefPubMedGoogle Scholar
  21. 21.
    Fearon ER, Vogelstein B. A genetic model for colorectal tumorigenesis. Cell. 1990;61(5):759–67.CrossRefPubMedGoogle Scholar
  22. 22.
    Allegra CJ, Paik S, Colangelo LH, Parr AL, Kirsch I, Kim G, et al. Prognostic value of thymidylate synthase, Ki-67, and p53 in patients with Dukes’ B and C colon cancer: a National Cancer Institute-National Surgical Adjuvant Breast and Bowel Project collaborative study. J Clin Oncol. 2003;21(2):241–50.CrossRefPubMedGoogle Scholar
  23. 23.
    Allegra CJ, Parr AL, Wold LE, Mahoney MR, Sargent DJ, Johnston P, et al. Investigation of the prognostic and predictive value of thymidylate synthase, p53, and Ki-67 in patients with locally advanced colon cancer. J Clin Oncol. 2002;20(7):1735–43.CrossRefPubMedGoogle Scholar
  24. 24.
    Belt EJ, Brosens RP, Delis-van Diemen PM, Bril H, Tijssen M, van Essen DF, et al. Cell cycle proteins predict recurrence in stage II and III colon cancer. Ann Surg Oncol. 2012;19(Suppl 3):S682–92.CrossRefPubMedGoogle Scholar
  25. 25.
    Demir L, Ekinci N, Erten C, Somali I, Can A, Dirican A, et al. The impact of cell proliferation markers and p53 mutation status on prognosis of non-metastatic colon cancer. J Surg Oncol. 2014;109(7):665–75.CrossRefPubMedGoogle Scholar
  26. 26.
    Flatmark K, Bjornland K, Johannessen HO, Hegstad E, Rosales R, Harklau L, et al. Immunomagnetic detection of micrometastatic cells in bone marrow of colorectal cancer patients. Clin Cancer Res. 2002;8(2):444–9.PubMedGoogle Scholar
  27. 27.
    Boye K, Nesland JM, Sandstad B, Haugland Haugen M, Maelandsmo GM, Flatmark K. EMMPRIN is associated with S100A4 and predicts patient outcome in colorectal cancer. Br J Cancer. 2012;107(4):667–74.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Flatmark K, Maelandsmo GM, Mikalsen SO, Nustad K, Varaas T, Rasmussen H, et al. Immunofluorometric assay for the metastasis-related protein S100A4: release of S100A4 from normal blood cells prohibits the use of S100A4 as a tumor marker in plasma and serum. Tumour Biol. 2004;25(1–2):31–40.CrossRefPubMedGoogle Scholar
  29. 29.
    Flatmark K, Pedersen KB, Nesland JM, Rasmussen H, Aamodt G, Mikalsen SO, et al. Nuclear localization of the metastasis-related protein S100A4 correlates with tumour stage in colorectal cancer. J Pathol. 2003;200(5):589–95.CrossRefPubMedGoogle Scholar
  30. 30.
    Do K, Wilsker D, Ji J, Zlott J, Freshwater T, Kinders RJ et al. Phase I study of single-agent AZD1775 (MK-1775), a Wee1 kinase inhibitor, in patients with refractory solid tumors. J Clin Oncol 2015;33(30):3409–15.Google Scholar
  31. 31.
    Guertin AD, Li J, Liu Y, Hurd MS, Schuller AG, Long B, et al. Preclinical evaluation of the WEE1 inhibitor MK-1775 as single-agent anticancer therapy. Mol Cancer Ther. 2013;12(8):1442–52.CrossRefPubMedGoogle Scholar
  32. 32.
    Perry JA, Kornbluth S. Cdc25 and Wee1: analogous opposites? Cell Div. 2007;2:12.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Matsuo T, Yamaguchi S, Mitsui S, Emi A, Shimoda F, Okamura H. Control mechanism of the circadian clock for timing of cell division in vivo. Science. 2003;302(5643):255–9.CrossRefPubMedGoogle Scholar
  34. 34.
    Huisman SA, Oklejewicz M, Ahmadi AR, Tamanini F, Ijzermans JN, van der Horst GT, et al. Colorectal liver metastases with a disrupted circadian rhythm phase shift the peripheral clock in liver and kidney. Int J Cancer. 2015;136(5):1024–32.CrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2016

Authors and Affiliations

  • Eivind Valen Egeland
    • 1
  • Kjersti Flatmark
    • 1
    • 2
    • 3
  • Jahn M. Nesland
    • 3
    • 4
  • Vivi Ann Flørenes
    • 4
  • Gunhild M. Mælandsmo
    • 1
    • 5
  • Kjetil Boye
    • 1
    • 6
    Email author
  1. 1.Department of Tumor Biology, Institute for Cancer Research, Norwegian Radium Hospital, Oslo University HospitalOsloNorway
  2. 2.Department of Gastroenterological Surgery, Norwegian Radium HospitalOslo University HospitalOsloNorway
  3. 3.Medical FacultyUniversity of OsloOsloNorway
  4. 4.Department of Pathology, Norwegian Radium HospitalOslo University HospitalOsloNorway
  5. 5.Department of Pharmacy, Faculty of Health SciencesUniversity of TromsøTromsøNorway
  6. 6.Department of Oncology, Norwegian Radium HospitalOslo University HospitalOsloNorway

Personalised recommendations