Enrichment of nuclear S100A4 during G2/M in colorectal cancer cells: possible association with cyclin B1 and centrosomes
- 415 Downloads
S100A4 promotes metastasis in several types of cancer, but the involved molecular mechanisms are still incompletely described. The protein is associated with a wide variety of biological functions and it locates to different subcellular compartments, including nuclei, cytoplasm and extracellular space. Nuclear expression of S100A4 has been associated with more advanced disease stage as well as poor outcome in colorectal cancer (CRC). The present study was initiated to investigate the nuclear function of S100A4 and thereby unravel potential biological mechanisms linking nuclear expression to a more aggressive phenotype. CRC cell lines show heterogeneity in nuclear S100A4 expression and preliminary experiments revealed cells in G2/M to have increased nuclear accumulation compared to G1 and S cells, respectively. Synchronization experiments validated nuclear S100A4 expression to be most prominent in the G2/M phase, but manipulating nuclear levels of S100A4 using lentiviral modified cells failed to induce changes in cell cycle distribution and proliferation. Proximity ligation assay did, however, demonstrate proximity between S100A4 and cyclin B1 in vitro, while confocal microscopy showed S100A4 to localize to areas corresponding to centrosomes in mitotic cells prior to chromosome segregation. This might indicate a novel and uncharacterized function of the metastasis-associated protein in CRC cells.
KeywordsColorectal cancer S100A4 Nucleus Cell cycle Cyclin B1 Centrosomes
We would like to thank Stein Waagene for carrying out the animal experiments and Dr. Vivi Ann Flørenes for valuable feedback and discussion. The present work was kindly supported by the Norwegian Cancer Society (Grant No [#4218523581] to EVE).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- 4.Malashkevich VN, Varney KM, Garrett SC, Wilder PT, Knight D, Charpentier TH, Ramagopal UA, Almo SC, Weber DJ, Bresnick AR (2008) Structure of Ca2+-bound S100A4 and its interaction with peptides derived from nonmuscle myosin-IIA. Biochemistry 47(18):5111–5126. doi: 10.1021/bi702537s PubMedCentralCrossRefPubMedGoogle Scholar
- 6.Flatmark K, Pedersen KB, Nesland JM, Rasmussen H, Aamodt G, Mikalsen SO, Bjornland K, Fodstad O, Maelandsmo GM (2003) Nuclear localization of the metastasis-related protein S100A4 correlates with tumour stage in colorectal cancer. J Pathol 200(5):589–595. doi: 10.1002/path.1381 CrossRefPubMedGoogle Scholar
- 19.Brunton H, Goodarzi AA, Noon AT, Shrikhande A, Hansen RS, Jeggo PA, Shibata A (2011) Analysis of human syndromes with disordered chromatin reveals the impact of heterochromatin on the efficacy of ATM-dependent G2/M checkpoint arrest. Mol Cell Biol 31(19):4022–4035. doi: 10.1128/mcb.05289-11 PubMedCentralCrossRefPubMedGoogle Scholar
- 25.el Bahassi M, Myer DL, McKenney RJ, Hennigan RF, Stambrook PJ (2006) Priming phosphorylation of Chk2 by polo-like kinase 3 (Plk3) mediates its full activation by ATM and a downstream checkpoint in response to DNA damage. Mutat Res 596(1–2):166–176. doi: 10.1016/j.mrfmmm.2005.12.002 CrossRefGoogle Scholar
- 36.Flatmark K, Maelandsmo GM, Mikalsen SO, Nustad K, Varaas T, Rasmussen H, Meling GI, Fodstad O, Paus E (2004) 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 25(1–2):31–40. doi: 10.1159/000077721 CrossRefPubMedGoogle Scholar
- 43.Murzik U, Hemmerich P, Weidtkamp-Peters S, Ulbricht T, Bussen W, Hentschel J, von Eggeling F, Melle C (2008) Rad54B targeting to DNA double-strand break repair sites requires complex formation with S100A11. Mol Biol Cell 19(7):2926–2935. doi: 10.1091/mbc.E07-11-1167 PubMedCentralCrossRefPubMedGoogle Scholar
- 48.Berge G, Costea DE, Berg M, Rasmussen H, Grotterod I, Lothe RA, Maelandsmo GM, Flatmark K (2011) Coexpression and nuclear colocalization of metastasis-promoting protein S100A4 and p53 without mutual regulation in colorectal carcinoma. Amino Acids 41(4):875–884. doi: 10.1007/s00726-010-0514-6 CrossRefPubMedGoogle Scholar
- 52.Myklebust MP, Li Z, Tran TH, Rui H, Knudsen ES, Elsaleh H, Fluge O, Vonen B, Myrvold HE, Leh S, Tveit KM, Pestell RG, Dahl O (2012) Expression of cyclin D1a and D1b as predictive factors for treatment response in colorectal cancer. Br J Cancer 107(10):1684–1691. doi: 10.1038/bjc.2012.463 PubMedCentralCrossRefPubMedGoogle Scholar
- 53.Fluge O, Gravdal K, Carlsen E, Vonen B, Kjellevold K, Refsum S, Lilleng R, Eide TJ, Halvorsen TB, Tveit KM, Otte AP, Akslen LA, Dahl O (2009) Expression of EZH2 and Ki-67 in colorectal cancer and associations with treatment response and prognosis. Br J Cancer 101(8):1282–1289. doi: 10.1038/sj.bjc.6605333 PubMedCentralCrossRefPubMedGoogle Scholar