The response of phyllodes tumor of the breast to anticancer therapy: An in vitro and ex vivo study

Urbaniak,Alicja; Jousheghany,Fariba; Yuan,Youzhong; Piña‑Oviedo,Sergio; Huczyński,Adam; Delgado,Magdalena; Kieber‑Emmons,Thomas; Monzavi‑Karbassi,Behjatolah; Chambers,Timothy  C.

doi:10.3892/ol.2019.10823

The response of phyllodes tumor of the breast to anticancer therapy: An in vitro and ex vivo study

Authors:
- Alicja Urbaniak
- Fariba Jousheghany
- Youzhong Yuan
- Sergio Piña‑Oviedo
- Adam Huczyński
- Magdalena Delgado
- Thomas Kieber‑Emmons
- Behjatolah Monzavi‑Karbassi
- Timothy C. Chambers
View Affiliations

Affiliations: Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA, Department of Pathology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA, Department of Bioorganic Chemistry, Faculty of Chemistry, Adam Mickiewicz University, Umultowska 89b, 61‑614 Poznań, Poland
Published online on: September 6, 2019 https://doi.org/10.3892/ol.2019.10823
Pages: 5097-5106
Copyright: © Urbaniak et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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

Phyllodes tumors of the breast (PTB) are uncommon stromal‑epithelial neoplasms, with the main recommended treatment being surgical removal. However, even with adequate resection, the risk of recurrence in the malignant form remains as high as 40%, and there is no recognized consensus on the most effective drugs for PTB. In the present study, an ex vivo model of malignant phyllodes and derived primary cell cultures were used to evaluate the effectiveness of a panel of different drugs, including Bcl‑2/Bcl‑xL inhibitor ABT‑263, salinomycin (SAL), doxorubicin (DOX), paclitaxel (TAX), vincristine (VCR), colchicine (COL) and cisplatin (CIS). ABT‑263, SAL and DOX were highly effective towards phyllodes spindle cells when assessed in the ex vivo model, contributing to ~98% tumor cell death. Furthermore, ABT‑263 was highly selective for tumor cells in this system, and exhibited little toxic effect on adjacent normal epithelial cells. Furthermore, consistent with findings in the ex vivo model, ABT‑263 was significantly less toxic towards MCF 10A non‑tumorigenic breast epithelial cells compared with SAL and DOX. A conditional reprogramming strategy was subsequently used, involving Rho kinase inhibition, to successfully generate primary phyllodes tumor cells that could be cultured for several passages. The primary cells were sensitive to DOX with an IC50 of 0.40±0.07 µM in a standard viability assay and the preliminary results were obtained indicating sensitivity to ABT‑263 and SAL. The present study demonstrated the feasibility of using explants and primary cells for drug discovery, selectively targeting PTB cells.

View Figures

View References

1	Parker SJ and Harries SA: Phyllodes tumors. Postgrad Med J. 77:428–435. 2001. View Article : Google Scholar : PubMed/NCBI
2	Strode M, Khoury T, Mangieri C and Takabe K: Update on the diagnosis and management of malignant phyllodes tumors of the breast. Breast. 33:91–96. 2017. View Article : Google Scholar : PubMed/NCBI
3	Roberts N and Runk DM: Aggressive malignant phyllodes tumor. Int J Surg Case Rep. 8:161–165. 2015. View Article : Google Scholar
4	Mishra SP, Tiwary SK, Mishra M and Khanna AK: Phyllodes tumor of breast: A review article. ISRN Surg. 2013:3614692013. View Article : Google Scholar : PubMed/NCBI
5	Norat F, Dreant N, Riah Y and Lebreton E: Ann ital chir extraordinary case of malignant phylloid tumor of the breast: Surgical reconstruction treatment (Italian). 80:475–478. 2009.PubMed/NCBI
6	Chaney AW, Pollack A, McNeese MD, Zagars GK, Pisters PWT, Pollock RE and Hunt KK: Primary treatment of cystosarcoma phyllodes of the breast. Cancer. 89:1502–1511. 2000. View Article : Google Scholar : PubMed/NCBI
7	Matar N, Soumani A, Noun M, Chraibi T, Himmi A, el Mansouri A, Aderdour M and Bekkay M: Phyllodes tumors of the breast. Forty one cases. J Gynecol Obstet Biol Reprod (Paris). 26:32–36. 1997.PubMed/NCBI
8	Belkacémi Y, Bousquet G, Marsiglia H, Ray-Coquard I, Magné N, Malard Y, Lacroix M, Gutierrez C, Senkus E, Christie D, et al: Phyllodes tumor of the breast. Int J Radiat Oncol Biol Phys. 70:492–500. 2008. View Article : Google Scholar : PubMed/NCBI
9	Guillot E, Couturaud B, Reyal F, Curnier A, Ravinet J, Laé M, Bollet M, Pierga JY, Salmon R and Fitoussi A: Breast cancer study group of the institut curie: Management of phyllodes breast tumors. Breast J. 17:129–137. 2011. View Article : Google Scholar : PubMed/NCBI
10	Morales-Vasquez F, Gonzalez-Angulo AM, Broglio K, Lopez-Basave HN, Gallardo D, Hortobagyi GN and De La Garza JG: Adjuvant chemotherapy with doxorubicin and dacarbazine has no effect in recurrence-free survival of malignant phyllodes tumors of the breast. Breast J. 13:551–556. 2007. View Article : Google Scholar : PubMed/NCBI
11	Hashimoto K, Mimura H, Arai Y, Doi M, Kojima Y, Tsugawa K and Nakajima Y: Successful preoperative chemoembolization in the treatment of a giant malignant phyllodes tumor. Cardiovasc Intervent Radiol. 39:1070–1075. 2016. View Article : Google Scholar : PubMed/NCBI
12	Shoemaker RH: The NCI60 human tumour cell line anticancer drug screen. Nat Rev Cancer. 6:813–823. 2006. View Article : Google Scholar : PubMed/NCBI
13	Burdall SE, Hanby AM, Lansdown MR and Speirs V: Breast cancer cell lines: Friend or foe. Breast Cancer Res. 5:89–95. 2003. View Article : Google Scholar : PubMed/NCBI
14	Witkiewicz AK, Rivadeneira DB, Ertel A, Kline J, Hyslop T, Schwartz GF, Fortina P and Knudsen ES: Association of RB/p16-pathway perturbations with DCIS recurrence: Dependence on tumor versus tissue microenvironment. Am J Pathol. 179:1171–1178. 2011. View Article : Google Scholar : PubMed/NCBI
15	Vargo-Gogola T and Rosen JM: Modelling breast cancer: One size does not fit all. Nat Rev Cancer. 7:659–672. 2007. View Article : Google Scholar : PubMed/NCBI
16	Dean JL, McClendon AK, Hickey TE, Butler LM, Tilley WD, Witkiewicz AK and Knudsen ES: Therapeutic response to CDK4/6 inhibition in breast cancer defined by ex vivo analyses of human tumors. Cell Cycle. 11:2756–2761. 2012. View Article : Google Scholar : PubMed/NCBI
17	Majumder B, Baraneedharan U, Thiyagarajan S, Radhakrishnan P, Narasimhan H, Dhandapani M, Brijwani N, Pinto DD, Prasath A, Shanthappa BU, et al: Predicting clinical response to anticancer drugs using an ex vivo platform that captures tumour heterogeneity. Nature Commun. 6:61692015. View Article : Google Scholar
18	Nagourney RA: Ex vivo programmed cell death and the prediction of response to chemotherapy. Curr Treat Options Oncol. 7:103–110. 2006. View Article : Google Scholar : PubMed/NCBI
19	Kuilman T, Michaloglou C, Mooi WJ and Peeper DS: The essence of senescence. Genes Dev. 24:2463–2479. 2010. View Article : Google Scholar : PubMed/NCBI
20	Roig AI, Eskiocak U, Hight SK, Kim SB, Delgado O, Souza RF, Spechler SJ, Wright WE and Shay JW: Immortalized epithelial cells derived from human colon biopsies express stem cell markers and differentiate in vitro. Gastroenterology. 138:1012–1021. 2010. View Article : Google Scholar : PubMed/NCBI
21	Liu X, Ory V, Chapman S, Yuan H, Albanese C, Kallakury B, Timofeeva OA, Nealon C, Dakic A, Simic V, et al: ROCK inhibitor and feeder cells induce the conditional reprogramming of epithelial cells. Am J Pathol. 180:599–607. 2012. View Article : Google Scholar : PubMed/NCBI
22	Saenz FR, Ory V, AlOtaiby M, Rosenfield S, Furlong M, Cavalli LR, Johnson MD, Liu X, Schlegel R, Wellstein A and Riegel AT: Conditionally reprogrammed normal and transformed mouse mammary epithelial cells display a progenitor-cell-like phenotype. PLoS One. 9:976662014. View Article : Google Scholar
23	Brown DD, Dabbs DJ, Lee AV, McGuire KP, Ahrendt GM, Bhargava R, Davidson NE, Brufsky AM, Johnson RR, Oesterreich S and McAuliffe PF: Developing in vitro models of human ductal carcinoma in situ from primary tissue explants. Breast Cancer Res Treat. 153:311–321. 2015. View Article : Google Scholar : PubMed/NCBI
24	Moo TA, Sanford R, Dang C and Morrow M: Overview of breast cancer therapy. PET Clin. 13:339–354. 2018. View Article : Google Scholar : PubMed/NCBI
25	Steinmetz MO and Prota AE: Microtubule-targeting agents: Strategies to hijack the cytoskeleton. Trends Cell Biol. 28:776–792. 2018. View Article : Google Scholar : PubMed/NCBI
26	Billard C: BH3 mimetics: Status of the field and new developments. Mol Cancer Ther. 12:1691–700. 2013. View Article : Google Scholar : PubMed/NCBI
27	Antoszczak M, Urbaniak A, Delgado M, Maj E, Borgström B, Wietrzyk J, Huczyński A, Yuan Y, Chambers TC and Strand D: Biological activity of doubly modified salinomycin analogs - Evaluation in vitro and ex vivo. Eur J Med Chem. 156:510–523. 2018. View Article : Google Scholar : PubMed/NCBI
28	Antoszczak M, Popiel K, Stefańska J, Wietrzyk J, Maj E, Janczak J, Michalska G, Brzezinski B and Huczynski A: Synthesis, cytotoxicity and antibacterial activity of new esters of polyether antibiotic-alinomycin. Eur J Med Chem. 76:435–444. 2014. View Article : Google Scholar : PubMed/NCBI
29	Van der Kuip H, Mürdter TE, Sonnenberg M, McClellan M, Gutzeit S, Gerteis A, Simon W, Fritz P and Aulitzky WE: Short term culture of breast cancer tissues to study the activity of the anticancer drug taxol in an intact tumor environment. BMC Cancer. 6:862006. View Article : Google Scholar : PubMed/NCBI
30	Dirks WG, MacLeod RA, Nakamura Y, Kohara A, Reid Y, Milch H, Drexler HG and Mizusawa H: Cell line cross-contamination initiative: An interactive reference database of STR profiles covering common cancer cell lines. Int J Cancer. 126:303–304. 2010. View Article : Google Scholar : PubMed/NCBI
31	Alley MC, Scudiere DA, Monks A, Czerwinski M, Shoemaker RH and Boyd MR: Validation of an automated microculture tetrazolium assay (MTA) to assess growth and drug sensitivity of human tumor cell lines. Proc Am Assoc Cancer Res. 27:389–391. 1986.
32	Alley MC, Scudiere DA, Monks A, Hursey ML, Czerwinski MJ, Fine DL, Abbott BJ, Mayo JG, Shoemaker RH and Boyd MR: Feasibility of drug screening with panels of human tumor cell lines using a microculture tetrazolium assay. Cancer Res. 48:589–601. 1988.PubMed/NCBI
33	Yerushalmi R, Woods R, Ravdin PM, Hayes MM and Gelmon KA: Ki67 in breast cancer: Prognostic and predictive potential. Lancet Oncol. 11:174–183. 2010. View Article : Google Scholar : PubMed/NCBI
34	Lin CK, Tsai WC, Lin YC and Yu JC: Biomarkers distinguishing mammary fibroepithelial neoplasms: A tissue microarray study. Appl Immunohistochem Mol Morphol. 22:433–441. 2014. View Article : Google Scholar : PubMed/NCBI
35	Went P, Dirnhofer S, Schöpf D, Moch H and Spizzo G: Expression and prognostic significance of EpCAM. J Cancer Mol. 3:169–174. 2008.
36	Martowicz A, Spizzo G, Gastl G and Untergasser G: Phenotype-dependent effects of EpCAM expression on growth and invasion of human breast cancer cell lines. BMC Cancer. 12:5012012. View Article : Google Scholar : PubMed/NCBI
37	Lewko WM, Vaghmar R, Maleckar JR, Husseini S, Montgomery CA, Thurman GB and Oldham RK: Cultured breast cystosarcoma phylloides cells and applications to patient therapy. Breast Cancer Res Treat. 17:131–138. 1990. View Article : Google Scholar : PubMed/NCBI
38	Tibbetts LM, Poisson MH, Tibbetts LL and Cummings FJ: A human breast stromal sarcoma cell line with features of malignant cystosarcoma phyllodes. Cancer. 62:2176–2182. 1988. View Article : Google Scholar : PubMed/NCBI
39	Jordan MA and Wilson L: Microtubules as a target for anticancer drugs. Nat Rev Cancer. 4:253–265. 2004. View Article : Google Scholar : PubMed/NCBI
40	Wheate NJ, Walker S, Craig GE and Oun R: The status of platinum anticancer drugs in the clinic and in clinical trials. Dalton Trans. 39:8113–8127. 2010. View Article : Google Scholar : PubMed/NCBI
41	Tait SW and Green DR: Mitochondrial regulation of cell death. Cold Spring Harb Perspect Bio. 5(pii): a0087062013. View Article : Google Scholar
42	Mituś JW, Blecharz P, Walasek T, Reinfuss M, Jakubowicz J and Kulpa J: Treatment of patients with distant metastases from phyllodes tumor of the breast. World J Surg. 40:323–328. 2016. View Article : Google Scholar : PubMed/NCBI
43	Liu M, Yang S, Liu B, Guo L, Bao X, Liu B and Dong L: Giant malignant phyllodes tumor of the breast: A rare case report and literature review. Oncol Lett. 12:121–124. 2016. View Article : Google Scholar : PubMed/NCBI
44	Allen R, Nixon D, York M and Coleman J: Successful chemotherapy for cystosarcoma phyllodes in a young woman. Arch Intern Med. 145:1127–1128. 1985. View Article : Google Scholar : PubMed/NCBI
45	Burton GV, Hart LL, Leight GS, Iglehart JD, McCarty KS Jr and Cox EB: Cystosarcoma phyllodes effective therapy with cisplatin and etoposide chemotherapy. Cancer. 63:2088–2092. 1989. View Article : Google Scholar : PubMed/NCBI
46	Lim SZ, Ng CCY, Rajasegaran V, Guan P, Selvarajan S, Thike AA, Nasir NDBM, Koh VCY, Tan BKT, Ong KW, et al: Genomic profile of breast sarcomas: A comparison with malignant phyllodes tumours. Breast Cancer Res Treat. 174:365–373. 2019. View Article : Google Scholar : PubMed/NCBI
47	Nozad S, Sheehan CE, Gay LM, Elvin JA, Vergilio JA, Suh J, Ramkissoon S, Schrock AB, Hirshfield KM, Ali N, et al: Comprehensive genomic profiling of malignant phyllodes tumorsof the breast. Breast Cancer Res Treat. 162:597–602. 2017. View Article : Google Scholar : PubMed/NCBI
48	Moore T and Lee AH: Expression of CD34 and bcl-2 in phyllodes tumours, fibroadenomas and spindle cell lesions of the breast. Histopathology. 38:62–67. 2001. View Article : Google Scholar : PubMed/NCBI
49	Jardim DL, Conley A and Subbiah V: Comprehensive characterization of malignant phyllodes tumor by whole genomic and proteomic analysis: Biological implications for targeted therapy opportunities. Orphanet J Rare Dis. 8:1122013. View Article : Google Scholar : PubMed/NCBI