Inhibition of Cdk2 activity decreases Aurora-A kinase centrosomal localization and prevents centrosome amplification in breast cancer cells

Leontovich,Alexey  A.; Salisbury,Jeffrey  L.; Veroux,Massimiliano; Tallarita,Tiziano; Billadeau,Daniel; McCubrey,James; Ingle,James; Galanis,Evanthia; D'Assoro,Antonino  B.

doi:10.3892/or.2013.2313

Inhibition of Cdk2 activity decreases Aurora-A kinase centrosomal localization and prevents centrosome amplification in breast cancer cells

Authors:
- Alexey A. Leontovich
- Jeffrey L. Salisbury
- Massimiliano Veroux
- Tiziano Tallarita
- Daniel Billadeau
- James McCubrey
- James Ingle
- Evanthia Galanis
- Antonino B. D'Assoro
View Affiliations

Affiliations: Department of Biochemistry and Molecular Biology, Mayo Clinic, College of Medicine, Rochester, MN, USA, Department of Surgery, Transplantation and Advanced Technologies, Organ Transplant Unit and Vascular Surgery, University of Catania, Catania, Italy, Medical Oncology, Mayo Clinic, College of Medicine, Rochester, MN, USA, Brody School of Medicine at East Carolina University, Greenville, NC, USA
Published online on: February 27, 2013 https://doi.org/10.3892/or.2013.2313
Pages: 1785-1788

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Centrosome amplification plays a key role in the origin of chromosomal instability (CIN) during cancer development and progression. In this study, MCF-7 breast cancer cell lines harboring abrogated p53 function (vMCF-7DNp53) were employed to investigate the relationship between induction of genotoxic stress, activation of cyclin-A/Cdk2 and Aurora-A oncogenic signalings and development of centrosome amplification. Introduction of genotoxic stress in the vMCF-7DNp53 cell line by treatment with hydroxyurea (HU) induced centrosome amplification that was mechanistically linked to Aurora-A kinase activity. In cells carrying defective p53, the development of centrosome amplification also occurred following treatment with another DNA damaging agent, methotrexate. Importantly, we demonstrated that Aurora-A kinase-induced centrosome amplification was mediated by Cdk2 kinase since molecular inhibition of Cdk2 activity by SU9516 suppressed Aurora-A centrosomal localization and consequent centrosome amplification. In addition, we employed vMCF-7DRaf-1 cells that display high levels of endogenous cyclin-A and demonstrated that molecular targeting of Aurora-A by Alisertib reduces cyclin-A expression. Taken together, these findings demonstrate a novel positive feed-back loop between cyclin-A/Cdk2 and Aurora-A pathways in the development of centrosome amplification in breast cancer cells. They also provide the translational rationale for targeting ‘druggable cell cycle regulators’ as an innovative therapeutic strategy to inhibit centrosome amplification and CIN in breast tumors resistant to conventional chemotherapeutic drugs.

View Figures

View References

1	Lengauer C, Kinzler KW and Vogelstein B: Genetic instabilities in human cancers. Nature. 6712:643–649. 1998. View Article : Google Scholar : PubMed/NCBI
2	Loeb LA: A mutator phenotype in cancer. Cancer Res. 61:3230–3239. 2001.PubMed/NCBI
3	D’Assoro AB, Lingle WL and Salisbury JL: Centrosome amplification and the development of cancer. Oncogene. 21:6146–6153. 2002.
4	Jin S and Levine AJ: The p53 functional circuit. J Cell Sci. 114:4139–4140. 2001.PubMed/NCBI
5	D’Assoro AB, Busby R, Suino K, Delva E, Almodovar-Mercado GJ, Johnson H, Folk C, Farrugia DJ, Vasile V, Stivala F and Salisbury JL: Genotoxic stress leads to centrosome amplification in breast cancer cell lines that have an inactive G1/S cell cycle checkpoint. Oncogene. 23:4068–4075. 2004.PubMed/NCBI
6	D’Assoro AB, Busby R, Acu ID, Quatraro C, Reinholz MM, Farrugia DJ, Schroeder MA, Allen C, Stivala F, Galanis E and Salisbury JL: Impaired p53 function leads to centrosome amplification, acquired ERalpha phenotypic heterogeneity and distant metastases in breast cancer MCF-7 xenografts. Oncogene. 27:3901–3911. 2008.PubMed/NCBI
7	Brinkley BR: Managing the centrosome numbers game: from chaos to stability in cancer cell division. Trends Cell Biol. 11:18–21. 2001. View Article : Google Scholar : PubMed/NCBI
8	D’Assoro AB, Barrett SL, Folk C, Negron VC, Boeneman K, Busby R, Whitehead C, Stivala F, Lingle WL and Salisbury JL: Amplified centrosomes in breast cancer: a potential indicator of tumor aggressiveness. Breast Cancer Res Treat. 75:25–34. 2002.PubMed/NCBI
9	Lingle WL, Lutz WH, Ingle JN, Maihle NJ and Salisbury JL: Centrosome hypertrophy in human breast tumors: implications for genomic stability and cell polarity. Proc Natl Acad Sci USA. 95:2950–2955. 1998. View Article : Google Scholar : PubMed/NCBI
10	Lingle WL, Barrett SL, Negron VC, D’Assoro AB, Boeneman K, Liu W, Whitehead CM, Reynolds C and Salisbury JL: Centrosome amplification drives chromosomal instability in breast tumor development. Proc Natl Acad Sci USA. 99:1978–1983. 2002. View Article : Google Scholar : PubMed/NCBI
11	Tarapore P and Fukasawa K: Loss of p53 and centrosome hyperamplification. Oncogene. 21:6234–6240. 2002. View Article : Google Scholar : PubMed/NCBI
12	Meraldi P, Lukas J, Fry AM, Bartek J and Nigg EA: Centrosome duplication in mammalian somatic cells requires E2F and Cdk2-cyclin A. Nat Cell Biol. 1:88–93. 1999. View Article : Google Scholar : PubMed/NCBI
13	Okuda M: The role of nucleophosmin in centrosome duplication. Oncogene. 21:6170–6174. 2002. View Article : Google Scholar : PubMed/NCBI
14	Mussman JG, Horn HF, Carroll PE, Okuda M, Tarapore P, Donehower LA and Fukasawa K: Synergistic induction of centrosome hyperamplification by loss of p53 and cyclin E overexpression. Oncogene. 19:1635–1646. 2000. View Article : Google Scholar : PubMed/NCBI
15	Nikonova AS, Astsaturov I, Serebriiskii IG, Dunbrack RL Jr and Golemis EA: Aurora A kinase (AURKA) in normal and pathological cell division. Cell Mol Life Sci. Aug 3–2012.(Epub ahead of print).
16	Li JJ, Weroha SJ, Lingle WL, Papa D, Salisbury JL and Li SA: Estrogen mediates Aurora-A overexpression, centrosome amplification, chromosomal instability, and breast cancer in female ACI rats. Proc Natl Acad Sci USA. 101:18123–18128. 2004. View Article : Google Scholar : PubMed/NCBI
17	Staff S, Isola J, Jumppanen M and Tanner M: Aurora-A gene is frequently amplified in basal-like breast cancer. Oncol Rep. 23:307–312. 2010.PubMed/NCBI
18	Yang H, He L, Kruk P, Nicosia SV and Cheng JQ: Aurora-A induces cell survival and chemoresistance by activation of Akt through a p53-dependent manner in ovarian cancer cells. Int J Cancer. 119:2304–2312. 2006. View Article : Google Scholar : PubMed/NCBI
19	D’Assoro AB, Leontovich A, Amato A, Ayers-Ringler JR, Quatraro C, Hafner K, Jenkins RB, Libra M, Ingle J, Stivala F, Galanis E and Salisbury JL: Abrogation of p53 function leads to metastatic transcriptome networks that typify tumor progression in human breast cancer xenografts. Int J Oncol. 37:1167–1176. 2010.PubMed/NCBI
20	Leontovich AA, Zhang S, Quatraro C, Iankov I, Veroux PF, Gambino MW, Degnim A, McCubrey J, Ingle J, Galanis E and D’Assoro AB: Raf-1 oncogenic signaling is linked to activation of mesenchymal to epithelial transition pathway in metastatic breast cancer cells. Int J Oncol. 40:1858–1864. 2012.PubMed/NCBI
21	Pinto AE, André S, Mendonça E, Silva G and Soares J: Overall survival in advanced breast cancer: relevance of progesterone receptor expression and DNA ploidy in fine-needle aspirates of 392 patients. Int J Biol Markers. 18:7–12. 2003.PubMed/NCBI
22	Duensing S and Münger K: Centrosomes, genomic instability, and cervical carcinogenesis. Crit Rev Eukaryot Gene. 13:9–23. 2003. View Article : Google Scholar
23	Levine AJ, Momand J and Finlay CA: The p53 tumour suppressor gene. Nature. 351:453–456. 1991. View Article : Google Scholar : PubMed/NCBI
24	Kronenwett U, Castro J, Roblick UJ, Fujioka K, Ostring C, Faridmoghaddam F, Laytragoon-Lewin N, Tribukait B and Auer G: Expression of cyclins A, E and topoisomerase II alpha correlates with centrosome amplification and genomic instability and influences the reliability of cytometric S-phase determination. BMC Cell Biol. 4:82003. View Article : Google Scholar : PubMed/NCBI
25	Lukasiewicz KB, Greenwood TM, Negron VC, Bruzek AK, Salisbury JL and Lingle WL: Control of centrin stability by Aurora A. PLoS One. 6:e212912011. View Article : Google Scholar : PubMed/NCBI