Inhibition of Cdk2 kinase activity selectively targets the CD44+/CD24-/Low stem-like subpopulation and restores chemosensitivity of SUM149PT triple-negative breast cancer cells

Opyrchal,Mateusz; Salisbury,Jeffrey  L.; Iankov,Ianko; Goetz,Mathew  P.; McCubrey,James; Gambino,Mario  W.; Malatino,Lorenzo; Puccia,Giuseppe; Ingle,James  N.; Galanis,Evanthia; D'Assoro,Antonino  B.

doi:10.3892/ijo.2014.2523

Inhibition of Cdk2 kinase activity selectively targets the CD44+/CD24-/Low stem-like subpopulation and restores chemosensitivity of SUM149PT triple-negative breast cancer cells

Authors:
- Mateusz Opyrchal
- Jeffrey L. Salisbury
- Ianko Iankov
- Mathew P. Goetz
- James McCubrey
- Mario W. Gambino
- Lorenzo Malatino
- Giuseppe Puccia
- James N. Ingle
- Evanthia Galanis
- Antonino B. D'Assoro
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Affiliations: Department of Medical Oncology, Mayo Clinic College of Medicine, Rochester, MN, USA, Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Rochester, MN, USA, Department of Microbiology and Immunology, East Carolina University, Greenville, NC, USA, Department of Internal Medicine, University of Catania, Catania, Italy
Published online on: June 25, 2014 https://doi.org/10.3892/ijo.2014.2523
Pages: 1193-1199

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Abstract

Inflammatory breast cancer (IBC) is an angioinvasive and most aggressive type of advanced breast cancer characterized by rapid proliferation, chemoresistance, early metastatic development and poor prognosis. IBC tumors display a triple-negative breast cancer (TNBC) phenotype characterized by centrosome amplification, high grade of chromosomal instability (CIN) and low levels of expression of estrogen receptor α (ERα), progesterone receptor (PR) and HER-2 tyrosine kinase receptor. Since the TNBC cells lack these receptors necessary to promote tumor growth, common treatments such as endocrine therapy and molecular targeting of HER-2 receptor are ineffective for this subtype of breast cancer. To date, not a single targeted therapy has been approved for non-inflammatory and inflammatory TNBC tumors and combination of conventional cytotoxic chemotherapeutic agents remains the standard therapy. IBC tumors generally display activation of epithelial to mesenchymal transition (EMT) that is functionally linked to a CD44+/CD24-/Low stem-like phenotype. Development of EMT and consequent activation of stemness programming is responsible for invasion, tumor self-renewal and drug resistance leading to breast cancer progression, distant metastases and poor prognosis. In this study, we employed the luminal ER+ MCF-7 and the IBC SUM149PT breast cancer cell lines to establish the extent to which high grade of CIN and chemoresistance were mechanistically linked to the enrichment of CD44+/CD24low/- CSCs. Here, we demonstrate that SUM149PT cells displayed higher CIN than MCF-7 cells characterized by higher percentage of structural and numerical chromosomal aberrations. Moreover, centrosome amplification, cyclin E overexpression and phosphorylation of retinoblastoma (Rb) were restricted to the stem-like CD44+/CD24-/Low subpopulation isolated from SUM149PT cells. Significantly, CD44+/CD24-/Low CSCs displayed resistance to conventional chemotherapy but higher sensitivity to SU9516, a specific cyclin-dependent kinase 2 (Cdk2) inhibitor, demonstrating that aberrant activation of cyclin E/Cdk2 oncogenic signaling is essential for the maintenance and expansion of CD44+/CD24-/Low CSC subpopulation in IBC. In conclusion, our findings propose a novel therapeutic approach to restore chemosensitivity and delay recurrence of IBC tumors based on the combination of conventional chemotherapy with small molecule inhibitors of the Cdk2 cell cycle kinase.

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1	Fernandez SV, Robertson FM, Pei J, Aburto-Chumpitaz L, Mu Z, Chu K, Alpaugh RK, Huang Y, Cao Y, Ye Z, Cai KQ, Boley KM, Klein-Szanto AJ, Devarajan K, Addya S and Cristofanilli M: Inflammatory breast cancer (IBC): clues for targeted therapies. Breast Cancer Res Treat. 140:23–33. 2013. View Article : Google Scholar : PubMed/NCBI
2	Yasumura K, Ogawa K, Ishikawa H, Takeshita T, Nakagawa Y and Osamura RY: Inflammatory carcinoma of the breast: characteristic findings of MR imaging. Breast Cancer. 4:161–169. 1997. View Article : Google Scholar : PubMed/NCBI
3	Yamauchi H, Woodward WA, Valero V, Alvarez RH, Lucci A, Buchholz TA, Iwamoto T, Krishnamurthy S, Yang W, Reuben JM, Hortobágyi GN and Ueno NT: Inflammatory breast cancer: what we know and what we need to learn. Oncologist. 17:891–899. 2012. View Article : Google Scholar : PubMed/NCBI
4	Liedtke C, Bernemann C, Kiesel L and Rody A: Genomic profiling in triple-negative breast cancer. Breast Care (Basel). 8:408–413. 2013. View Article : Google Scholar : PubMed/NCBI
5	Qiu M, Peng Q, Jiang I, Carroll C, Han G, Rymer I, Lippincott J, Zachwieja J, Gajiwala K, Kraynov E, Thibault S, Stone D, Gao Y, Sofia S, Gallo J, Li G, Yang J, Li K and Wei P: Specific inhibition of Notch1 signaling enhances the antitumor efficacy of chemotherapy in triple negative breast cancer through reduction of cancer stem cells. Cancer Lett. 328:261–270. 2013. View Article : Google Scholar : PubMed/NCBI
6	Cortazar P, Zhang L, Untch M, Mehta K, Costantino JP, Wolmark N, Bonnefoi H, Cameron D, Gianni L, Valagussa P, Swain SM, Prowell T, Loibl S, Wickerham DL, Bogaerts J, Baselga J, Perou C, Blumenthal G, Blohmer J, Mamounas EP, Bergh J, Semiglazov V, Justice R, Eidtmann H, Paik S, Piccart M, Sridhara R, Fasching PA, Slaets L, Tang S, Gerber B, Geyer CE Jr, Pazdur R, Ditsch N, Rastogi P, Eiermann W and von Minckwitz G: Pathological complete response and long-term clinical benefit in breast cancer: the CTNeoBC pooled analysis. Lancet. Feb 14–2014.(Epub ahead of print).
7	Crown J, O’Shaughnessy J and Gullo G: Emerging targeted therapies in triple-negative breast cancer. Ann Oncol. 23(Suppl 6): vi56–vi65. 2012. View Article : Google Scholar : PubMed/NCBI
8	Greenup R, Buchanan A, Lorizio W, Rhoads K, Chan S, Leedom T, King R, McLennan J, Crawford B, Kelly Marcom P and Shelley Hwang E: Prevalence of BRCA mutations among women with triple-negative breast cancer (TNBC) in a genetic counseling cohort. Ann Surg Oncol. 20:3254–3258. 2013. View Article : Google Scholar : PubMed/NCBI
9	Venkitaraman AR: Cancer susceptibility and the functions of BRCA1 and BRCA2. Cell. 108:171–182. 2002. View Article : Google Scholar : PubMed/NCBI
10	Farrugia DJ, Agarwal MK, Pankratz VS, Deffenbaugh AM, Pruss D, Frye C, Wadum L, Johnson K, Mentlick J, Tavtigian SV, Goldgar DE and Couch FJ: Functional assays for classification of BRCA2 variants of uncertain significance. Cancer Res. 68:3523–3531. 2008. View Article : Google Scholar : PubMed/NCBI
11	Kais Z, Chiba N, Ishioka C and Parvin JD: Functional differences among BRCA1 missense mutations in the control of centrosome duplication. Oncogene. 31:799–804. 2012. View Article : Google Scholar : PubMed/NCBI
12	Kolupaeva V and Basilico C: Overexpression of cyclin E/CDK2 complexes overcomes FGF-induced cell cycle arrest in the presence of hypophosphorylated Rb proteins. Cell Cycle. 11:2557–2566. 2012. View Article : Google Scholar : PubMed/NCBI
13	Reed SI: Control of the G1/S transition. Cancer Surv. 29:7–23. 1997.
14	D’Assoro AB, Lingle WL and Salisbury JL: Centrosome amplification and the development of cancer. Oncogene. 21:6146–6153. 2002.
15	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
16	Hanashiro K, Kanai M, Geng Y, Sicinski P and Fukasawa K: Roles of cyclins A and E in induction of centrosome amplification in p53-compromised cells. Oncogene. 27:5288–5302. 2008. View Article : Google Scholar : PubMed/NCBI
17	Mani SA, Guo W, Liao MJ, Eaton EN, Ayyanan A, Zhou AY, Brooks M, Reinhard F, Zhang CC, Shipitsin M, Campbell LL, Polyak K, Brisken C, Yang J and Weinberg RA: The epithelial mesenchymal transition generates cells with properties of stem cells. Cell. 133:704–715. 2008. View Article : Google Scholar
18	Hwang-Verslues WW, Kuo WH, Chang PH, Pan CC, Wang HH, Tsai ST, Jeng YM, Shew JY, Kung JT, Chen CH, Lee EY, Chang KJ and Lee WH: Multiple lineages of human breast cancer stem/progenitor cells identified by profiling with stem cell markers. PLoS One. 4:83772009. View Article : Google Scholar : PubMed/NCBI
19	Shipitsin M, Campbell LL, Argani P, Weremowicz S, Bloushtain-Qimron N, Yao J, Nikolskaya T, Serebryiskaya T, Beroukhim R, Hu M, Halushka MK, Sukumar S, Parker LM, Anderson KS, Harris LN, Garber JE, Richardson AL, Schnitt SJ, Nikolsky Y, Gelman RS and Polyak K: Molecular definition of breast tumor heterogeneity. Cancer Cell. 11:259–273. 2007. View Article : Google Scholar : PubMed/NCBI
20	D’Assoro AB, Liu T, Quatraro C, Amato A, Opyrchal M, Leontovich A, Ikeda Y, Ohmine S, Lingle W, Suman V, Ecsedy J, Iankov I, Di Leonardo A, Ayers-Inglers J, Degnim A, Billadeau D, McCubrey J, Ingle J, Salisbury JL and Galanis E: The mitotic kinase Aurora-A promotes distant metastases by inducing epithelial-to-mesenchymal transition in ERα(+) breast cancer cells. Oncogene. 33:599–610. 2014.PubMed/NCBI
21	Lee HE, Kim JH, Kim YJ, Choi SY, Kim SW, Kang E, Chung IY, Kim IA, Kim EJ, Choi Y, Ryu HS and Park SY: An increase in cancer stem cell population after primary systemic therapy is a poor prognostic factor in breast cancer. Br J Cancer. 104:1730–1738. 2011. View Article : Google Scholar : PubMed/NCBI
22	Arima Y, Hayashi N, Hayashi H, Sasaki M, Kai K, Sugihara E, Abe E, Yoshida A, Mikami S, Nakamura S and Saya H: Loss of p16 expression is associated with the stem cell characteristics of surface markers and therapeutic resistance in estrogen receptor-negative breast cancer. Int J Cancer. 130:2568–2579. 2012. View Article : Google Scholar : PubMed/NCBI
23	Prat A, Karginova O, Parker JS, Fan C, He X, Bixby L, Harrell JC, Roman E, Adamo B, Troester M and Perou CM: Characterization of cell lines derived from breast cancers and normal mammary tissues for the study of the intrinsic molecular subtypes. Breast Cancer Res Treat. 142:237–255. 2013. View Article : Google Scholar : PubMed/NCBI
24	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
25	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
26	Iovino F, Lentini L, Amato A and Di Leonardo A: RB acute loss induces centrosome amplification and aneuploidy in murine primary fibroblasts. Mol Cancer. 5:382006. View Article : Google Scholar : PubMed/NCBI
27	Mayer IA, Abramson VG, Lehmann BD and Pietenpol JA: New strategies for triple-negative breast cancer - deciphering the heterogeneity. Clin Cancer Res. 20:782–790. 2014. View Article : Google Scholar : PubMed/NCBI
28	Mehta RS: Dose-dense and/or metronomic schedules of specific chemotherapies consolidate the chemosensitivity of triple-negative breast cancer: a step toward reversing triple-negative paradox. J Clin Oncol. 26:3286–3288. 2008. View Article : Google Scholar
29	Liedtke C, Mazouni C, Hess KR, André F, Tordai A, Mejia JA, Symmans WF, Gonzalez-Angulo AM, Hennessy B, Green M, Cristofanilli M, Hortobagyi GN and Pusztai L: Response to neoadjuvant therapy and long-term survival in patients with triple-negative breast cancer. J Clin Oncol. 26:1275–1281. 2008. View Article : Google Scholar : PubMed/NCBI
30	Cristofanilli M, Buzdar AU, Sneige N, Smith T, Wasaff B, Ibrahim N, Booser D, Rivera E, Murray JL, Valero V, Ueno N, Singletary ES, Hunt K, Strom E, McNeese M, Stelling C and Hortobagyi GN: Paclitaxel in the multimodality treatment for inflammatory breast carcinoma. Cancer. 92:1775–1782. 2001. View Article : Google Scholar : PubMed/NCBI
31	Takahashi T, Akashi-Tanaka S, Fukutomi T, Watanabe T, Katsumata N, Miyakawa K, Hasegawa T and Tsuda H: Two special types of breast cancer presenting as progressive disease after neoadjuvant chemotherapy with docetaxel plus doxorubicin. Breast Cancer. 8:234–237. 2001. View Article : Google Scholar : PubMed/NCBI
32	Leopold PL, Vincent J and Wang H: A comparison of epithelial-to-mesenchymal transition and re-epithelialization. Semin Cancer Biol. 22:471–483. 2012. View Article : Google Scholar : PubMed/NCBI
33	Rosen EM and Pishvaian MJ: Targeting the BRCA1/2 tumor suppressors. Curr Drug Targets. 15:17–31. 2014. View Article : Google Scholar
34	Khoury-Haddad H, Guttmann-Raviv N, Ipenberg I, Huggins D, Jeyasekharan AD and Ayoub N: PARP1-dependent recruitment of KDM4D histone demethylase to DNA damage sites promotes double-strand break repair. Proc Natl Acad Sci USA. 111:E728–E737. 2014. View Article : Google Scholar : PubMed/NCBI
35	Węsierska-Gądek J, Zulehner N, Ferk F, Składanowski A, Komina O and Maurer M: PARP inhibition potentiates the cytotoxic activity of C-1305, a selective inhibitor of topoisomerase II, in human BRCA1-positive breast cancer cells. Biochem Pharmacol. 84:1318–1331. 2012.PubMed/NCBI
36	Shen Y, Rehman FL, Feng Y, Boshuizen J, Bajrami I, Elliott R, Wang B, Lord CJ, Post LE and Ashworth A: BMN 673, a novel and highly potent PARP1/2 inhibitor for the treatment of human cancers with DNA repair deficiency. Clin Cancer Res. 19:5003–5015. 2013. View Article : Google Scholar : PubMed/NCBI
37	Fojo T and Bates S: Mechanisms of resistance to PARP inhibitors - three and counting. Cancer Discov. 3:20–23. 2013. View Article : Google Scholar : PubMed/NCBI
38	Yin S, Xu L, Bandyopadhyay S, Sethi S and Reddy KB: Cisplatin and TRAIL enhance breast cancer stem cell death. Int J Oncol. 39:891–898. 2011.PubMed/NCBI
39	Potemski P, Kusińska R, Pasz-Walczak G, Piekarski JH, Watała C, Płuciennik E, Bednarek AK and Kordek R: Prognostic relevance of cyclin E expression in operable breast cancer. Med Sci Monit. 15:MT34–MT40. 2009.PubMed/NCBI
40	Willmarth NE, Albertson DG and Ethier SP: Chromosomal instability and lack of cyclin E regulation in hCdc4 mutant human breast cancer cells. Breast Cancer Res. 6:R531–R539. 2004. View Article : Google Scholar : PubMed/NCBI