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Open Access 01-12-2016 | Research article

ANLN is a prognostic biomarker independent of Ki-67 and essential for cell cycle progression in primary breast cancer

Authors: Kristina Magnusson, Gabriela Gremel, Lisa Rydén, Victor Pontén, Mathias Uhlén, Anna Dimberg, Karin Jirström, Fredrik Pontén

Published in: BMC Cancer | Issue 1/2016

Abstract

Background

Anillin (ANLN), an actin-binding protein required for cytokinesis, has recently been presented as part of a prognostic marker panel in breast cancer. The objective of the current study was to further explore the prognostic and functional value of ANLN as a single biomarker in breast cancer.

Methods

Immunohistochemical assessment of ANLN protein expression was performed in two well characterized breast cancer cohorts (n = 484) with long-term clinical follow-up data and the results were further validated at the mRNA level in a publicly available transcriptomics dataset. The functional relevance of ANLN was investigated in two breast cancer cell lines using RNA interference.

Results

High nuclear fraction of ANLN in breast tumor cells was significantly associated with large tumor size, high histological grade, high proliferation rate, hormone receptor negative tumors and poor prognosis in both examined cohorts. Multivariable analysis showed that the association between ANLN and survival was significantly independent of age in cohort I and significantly independent of proliferation, as assessed by Ki-67 expression in tumor cells, age, tumor size, ER and PR status, HER2 status and nodal status in cohort II. Analysis of ANLN mRNA expression confirmed that high expression of ANLN was significantly correlated to poor overall survival in breast cancer patients. Consistent with the role of ANLN during cytokinesis, transient knock-down of ANLN protein expression in breast cancer cell lines resulted in an increase of senescent cells and an accumulation of cells in the G2/M phase of the cell cycle with altered cell morphology including large, poly-nucleated cells. Moreover, ANLN siRNA knockdown also resulted in decreased expression of cyclins D1, A2 and B1.

Conclusions

ANLN expression in breast cancer cells plays an important role during cell division and a high fraction of nuclear ANLN expression in tumor cells is correlated to poor prognosis in breast cancer patients, independent of Ki-67, tumor size, hormone receptor status, HER2 status, nodal status and age.

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IARC. Cancer incidence in five continents. Volume IX. IARC Sci Publ. No 160. Lyon: IARC; 2008. p. 1–837.

Brohede J. Cancer incidence in Sweden 2013. In: Book cancer incidence in Sweden 2013. 2014.

Oegema K, Savoian MS, Mitchison TJ, Field CM. Functional analysis of a human homologue of the drosophila actin binding protein anillin suggests a role in cytokinesis. J Cell Biol. 2000;150:539–52.CrossRefPubMedPubMedCentral

Piekny AJ, Glotzer M. Anillin is a scaffold protein that links RhoA, actin, and myosin during cytokinesis. Curr Biol. 2008;18:30–6.CrossRefPubMed

Hickson GR, O’Farrell PH. Rho-dependent control of anillin behavior during cytokinesis. J Cell Biol. 2008;180:285–94.CrossRefPubMedPubMedCentral

Suzuki C, Daigo Y, Ishikawa N, Kato T, Hayama S, Ito T, Tsuchiya E, Nakamura Y. ANLN plays a critical role in human lung carcinogenesis through the activation of RHOA and by involvement in the phosphoinositide 3-kinase/AKT pathway. Cancer Res. 2005;65:11314–25.CrossRefPubMed

Piekny AJ, Maddox AS. The myriad roles of Anillin during cytokinesis. Semin Cell Dev Biol. 2010;21:881–91.CrossRefPubMed

Hall PA, Todd CB, Hyland PL, McDade SS, Grabsch H, Dattani M, Hillan KJ, Russell SE. The septin-binding protein anillin is overexpressed in diverse human tumors. Clin Cancer Res. 2005;11:6780–6.CrossRefPubMed

Olakowski M, Tyszkiewicz T, Jarzab M, Krol R, Oczko-Wojciechowska M, Kowalska M, Kowal M, Gala GM, Kajor M, Lange D, et al. NBL1 and anillin (ANLN) genes over-expression in pancreatic carcinoma. Folia Histochem Cytobiol. 2009;47:249–55.CrossRefPubMed

10.

Shimizu S, Seki N, Sugimoto T, Horiguchi S, Tanzawa H, Hanazawa T, Okamoto Y. Identification of molecular targets in head and neck squamous cell carcinomas based on genome-wide gene expression profiling. Oncol Rep. 2007;18:1489–97.PubMed

11.

Skrzypski M, Jassem E, Taron M, Sanchez JJ, Mendez P, Rzyman W, Gulida G, Raz D, Jablons D, Provencio M, et al. Three-gene expression signature predicts survival in early-stage squamous cell carcinoma of the lung. Clin Cancer Res. 2008;14:4794–9.CrossRefPubMed

12.

Ronkainen H, Hirvikoski P, Kauppila S, Vaarala MH. Anillin expression is a marker of favourable prognosis in patients with renal cell carcinoma. Oncol Rep. 2011;25:129–33.PubMed

13.

Leary PC O, Penny SA, Dolan RT, Kelly CM, Madden SF, Rexhepaj E, Brennan DJ, McCann AH, Ponten F, Uhlen M, et al. Systematic antibody generation and validation via tissue microarray technology leading to identification of a novel protein prognostic panel in breast cancer. BMC Cancer. 2013;13:175.CrossRef

14.

Zhou W, Wang Z, Shen N, Pi W, Jiang W, Huang J, Hu Y, Li X, Sun L. Knockdown of ANLN by lentivirus inhibits cell growth and migration in human breast cancer. Mol Cell Biochem. 2015;398:11–9.CrossRefPubMed

15.

Kampf C, Olsson I, Ryberg U, Sjostedt E, Ponten F. Production of tissue microarrays, immunohistochemistry staining and digitalization within the human protein atlas. J Vis Exp. 2012;(63):1–8.

16.

Elkabets M, Gifford AM, Scheel C, Nilsson B, Reinhardt F, Bray MA, Carpenter AE, Jirstrom K, Magnusson K, Ebert BL, et al. Human tumors instigate granulin-expressing hematopoietic cells that promote malignancy by activating stromal fibroblasts in mice. J Clin Invest. 2011;121:784–99.CrossRefPubMedPubMedCentral

17.

Svensson KJ, Christianson HC, Kucharzewska P, Fagerstrom V, Lundstedt L, Borgquist S, Jirstrom K, Belting M. Chondroitin sulfate expression predicts poor outcome in breast cancer. Int J Oncol. 2011;39:1421–8.PubMed

18.

Ryden L, Jonsson PE, Chebil G, Dufmats M, Ferno M, Jirstrom K, Kallstrom AC, Landberg G, Stal O, Thorstenson S, Nordenskjold B. Two years of adjuvant tamoxifen in premenopausal patients with breast cancer: a randomised, controlled trial with long-term follow-up. Eur J Cancer. 2005;41:256–64.CrossRefPubMed

19.

Brennan DJ, Laursen H, O’Connor DP, Borgquist S, Uhlen M, Gallagher WM, Ponten F, Millikan RC, Ryden L, Jirstrom K. Tumor-specific HMG-CoA reductase expression in primary premenopausal breast cancer predicts response to tamoxifen. Breast Cancer Res. 2011;13:R12.CrossRefPubMedPubMedCentral

20.

Jirstrom K, Ryden L, Anagnostaki L, Nordenskjold B, Stal O, Thorstenson S, Chebil G, Jonsson PE, Ferno M, Landberg G. Pathology parameters and adjuvant tamoxifen response in a randomised premenopausal breast cancer trial. J Clin Pathol. 2005;58:1135–42.CrossRefPubMedPubMedCentral

21.

Stendahl M, Ryden L, Nordenskjold B, Jonsson PE, Landberg G, Jirstrom K. High progesterone receptor expression correlates to the effect of adjuvant tamoxifen in premenopausal breast cancer patients. Clin Cancer Res. 2006;12:4614–8.CrossRefPubMed

22.

Ponten F, Schwenk JM, Asplund A, Edqvist PH. The Human Protein Atlas as a proteomic resource for biomarker discovery. J Intern Med. 2011;270:428–46.

23.

Hjelm B, Forsstrom B, Igel U, Johannesson H, Stadler C, Lundberg E, Ponten F, Sjoberg A, Rockberg J, Schwenk JM, et al. Generation of monospecific antibodies based on affinity capture of polyclonal antibodies. Protein Sci. 2011;20:1824–35.CrossRefPubMedPubMedCentral

24.

Cole MP, Jones CT, Todd ID. A new anti-oestrogenic agent in late breast cancer. An early clinical appraisal of ICI46474. Br J Cancer. 1971;25:270–5.CrossRefPubMedPubMedCentral

25.

Rivera-Guevara C, Camacho J. Tamoxifen and its new derivatives in cancer research. Recent Pat Anticancer Drug Discov. 2011;6:237–45.CrossRefPubMed

26.

Early Breast Cancer Trialists’ Collaborative Group. Tamoxifen for early breast cancer: an overview of the randomised trials. Lancet. 1998;351:1451–67.

27.

Gnant M, Harbeck N, Thomssen C. St. Gallen 2011: summary of the consensus discussion. Breast Care (Basel). 2011;6:136–41.CrossRef

28.

Chanrion M, Negre V, Fontaine H, Salvetat N, Bibeau F, Mac Grogan G, Mauriac L, Katsaros D, Molina F, Theillet C, Darbon JM. A gene expression signature that can predict the recurrence of tamoxifen-treated primary breast cancer. Clin Cancer Res. 2008;14:1744–52.CrossRefPubMedPubMedCentral

29.

Sinicropi D, Qu K, Collin F, Crager M, Liu ML, Pelham RJ, Pho M, Dei Rossi A, Jeong J, Scott A, et al. Whole transcriptome RNA-Seq analysis of breast cancer recurrence risk using formalin-fixed paraffin-embedded tumor tissue. PLoS One. 2012;7:e40092.CrossRefPubMedPubMedCentral

30.

Brennan DJ, O’Connor DP, Rexhepaj E, Ponten F, Gallagher WM. Antibody-based proteomics: fast-tracking molecular diagnostics in oncology. Nat Rev Cancer. 2010;10:605–17.CrossRefPubMed

31.

Coates AS, Winer EP, Goldhirsch A, Gelber RD, Gnant M, Piccart-Gebhart M, Thurlimann B, Senn HJ. Tailoring therapies-improving the management of early breast cancer: St Gallen International Expert Consensus on the Primary Therapy of Early Breast Cancer 2015. Ann Oncol. 2015;26:1533–46.CrossRefPubMedPubMedCentral

32.

Fagerberg L, Hallstrom BM, Oksvold P, Kampf C, Djureinovic D, Odeberg J, Habuka M, Tahmasebpoor S, Danielsson A, Edlund K, et al. Analysis of the human tissue-specific expression by genome-wide integration of transcriptomics and antibody-based proteomics. Mol Cell Proteomics. 2014;13:397–406.CrossRefPubMed

33.

Uhlen M, Fagerberg L, Hallstrom BM, Lindskog C, Oksvold P, Mardinoglu A, Sivertsson A, Kampf C, Sjostedt E, Asplund A, et al. Proteomics. Tissue-based map of the human proteome. Science. 2015;347:1260419.CrossRefPubMed

34.

Zhao W-m, Fang G. Anillin is a substrate of anaphase-promoting complex/cyclosome (APC/C) that controls spatial contractility of myosin during late cytokinesis. J Biol Chem. 2005;280:33516–24.CrossRefPubMed

35.

Straight AF, Field CM, Mitchison TJ. Anillin binds nonmuscle myosin II and regulates the contractile ring. Mol Biol Cell. 2005;16:193–201.CrossRefPubMedPubMedCentral

Title: ANLN is a prognostic biomarker independent of Ki-67 and essential for cell cycle progression in primary breast cancer
Authors: Kristina Magnusson
Gabriela Gremel
Lisa Rydén
Victor Pontén
Mathias Uhlén
Anna Dimberg
Karin Jirström
Fredrik Pontén
Publication date: 01-12-2016
Publisher: BioMed Central
Published in: BMC Cancer / Issue 1/2016
Electronic ISSN: 1471-2407
DOI: https://doi.org/10.1186/s12885-016-2923-8

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Abstract

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Methods

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