PROBING BREAST CANCER THERAPEUTIC RESPONSES BY DNA CONTENT PROFILING
Background. Discrepancies in the interpretation of breast cancer therapeutic responses still exist mainly because of lack of standardized assessment criteria and methods.
Objective. DNA content profiling of cells in the affected (cancerous) tissue before and after neoadjuvant chemotherapy (NAC) was applied to facilitate interpretation of therapeutic responses.
Methods. Both diagnostic biopsy and operation materials representing the tissue of primary tumors surgically removed after NAC were subjected to DNA image cytometry. Polyploidy and aneuploidy in DNA histograms were evaluated with a prognostic Auer typing. Stemline DNA index (DI) values and percentages of cells that polyploidize (>4.5C) were also determined. Immunofluorescence staining was applied to evaluate proliferation (Ki-67), invasiveness (CD44), and self-renewal factors characteristic for stem cells (SOX2 and NANOG).
Results. DNA content profiles of 12 breast cancer cases, of which 7 were triple-negative, revealed the features of tumor non-responsiveness to NAC in 7 cases, of which 5 were triple-negative. Among non-responsive cases there were 3 cases that showed enhanced polyploidization, suggesting the negative NAC effect. Near-triploid (DI=1.26-1.74) triple-negative cases were determined as most resistant to NAC. Cycling near-triploid cells may contribute to the excessive numbers of >4.5C cells. Polyploid cells were positive for Ki-67, CD44, SOX2, and NANOG.
Conclusions. DNA content profiling data provide additional helpful information for interpreting therapeutic responses in NAC-treated breast cancers. Polyploid tumor cells possessing stem cell features can be induced by NAC. Because NAC effects in some cases may be unfavorable, the use of the further treatment strategy should be carefully considered.
Ragaz J, Baird R, Rebbeck P, Goldie J, Coldman A, Spinelli J. Neoadjuvant (preoperative) chemotherapy for breast cancer. Cancer 1985; 56: 719−724.
Buchholz TA, Hunt KK, Whitman GJ, Sahin AA, Hortobagyi GN. Neoadjuvant chemotherapy for breast carcinoma: Multidisciplinary considerations of benefits and risks. Cancer 2003; 98: 1150−1160.
Sahoo S, Lester SC. Pathology of breast carcinomas after neoadjuvant chemotherapy: An overview with recommendations on specimen processing and reporting. Arch Pathol Lab Med 2009; 133: 633−642.
Horii R, Akiyama F. Histological assessment of therapeutic response in breast cancer. Breast Cancer 2016; 23: 540−545.
Sahoo S, Lester SC. Pathology considerations in patients treated with neoadjuvant chemotherapy. Surg Pathol Clin 2012; 5: 749−774.
Kuroi K, Toi M, Tsuda H, Kurosumi M, Akiyama F. Issues in the assessment of the pathologic effect of primary systemic therapy for breast cancer. Breast cancer 2006; 13: 38−48.
Weaver BAA, Cleveland DM. Does aneuploidy cause cancer? Curr Opin Cell Biol 2006; 18: 658−667.
Gordon DJ, Resio B, Pellman D. Causes and consequences of aneuploidy in cancer. Nat Rev Genet 2012; 13: 189−203.
Swanton C, Nicke B, Schuett M, Eklund AC, Ng C, Li Q, et al. Chromosomal instability determines taxane response. Proc Natl Acad Sci U S A 2009; 106: 8671−8676.
Martelotto LG, Ng CKY, Piscuoglio S, Weigelt B, Reis-Filho JS. Breast cancer intra-tumor heterogeneity. Breast Cancer Res 2014; 16: R48.
McGranahan N, Swanton C. Clonal heterogeneity and tumor evolution: Past, present, and the future. Cell 2017; 168: 613−628.
Gerashchenko BI, Huna A, Erenpreisa J. Characterization of breast cancer DNA content profiles as a prognostic tool. Exp Oncol 2014; 36: 219−225.
Coward J, Harding A. Size does matter: Why polyploid tumor cells are critical drug targets in the war on cancer. Front Oncol 2014; 4: Article 123.
Dayal JHS, Sales MJ, Corver WE, Purdie CA, Jordan LB, Quinlan PR, et al. Multiparameter DNA content analysis identifies distinct groups in primary breast cancer. Br J Cancer 2013; 108: 873−880.
Kim C, Gao R, Sei E, Brandt R, Hartman J, Hatschek T, et al. Chemoresistance evolution in triple-negative breast cancer delineated by single-cell sequencing. Cell 2018; 173: 879−893.
Auer GU, Caspersson TO, Wallgren AS. DNA content and survival in mammary carcinoma. Anal Quant Cytol 1980; 2: 161−165.
Ogston KN, Miller ID, Payne S, Hutcheon AW, Sarkar TK, Smith I, et al. A new histological grading system to assess response of breast cancers to primary chemotherapy: prognostic significance and survival. Breast 2003; 12: 320−327.
Erenpreisa J, Freivalds T. Anisotropic staining of apurinic acid with toluidine blue. Histochemistry 1979; 60: 321−325.
Gerashchenko BI, Salmina K, Eglitis J, Huna A, Grjunberga V, Erenpreisa J. Disentangling the aneuploidy and senescence paradoxes: A study of triploid breast cancers non-responsive to neoadjuvant therapy. Histochem Cell Biol 2016; 145: 497−508.
Haroske G, Baak JPA, Danielsen H, Giroud F, Gschwendtner A, Oberholzer M, et al. Fourth updated ESACP consensus report on diagnostic DNA image cytometry. Anal Cell Pathol 2001; 23: 89−95.
Barlogie B, Hittelman W, Spitzer G, Trujillo JM, Hart JS, Smallwood L, et al. Correlation of DNA distribution abnormalities with cytogenetic findings in human adult leukemia and lymphoma. Cancer Res 1977; 37: 4400−4407.
Fallenius AG, Auer GU, Carstensen JM. Prognostic significance of DNA measurements in 409 consecutive breast cancer patients. Cancer 1988; 62: 331−341.
Leonardi E, Cristofori A, Caffo O, Dalla Palma P. Cytometric DNA analysis and prognostic biomarkers in breast carcinoma. Expression of P53 product in the different ploidy classes. Anal Cell Pathol 1997; 15: 31−45.
Schulze S, Petersen I. Gender and ploidy in cancer survival. Cell Oncol 2011; 34: 199−208.
Erenpreisa J, Salmina K, Huna A, Kosmacek EA, Cragg MS, Ianzini F, et al. Polyploid tumour cells elicit paradiploid progeny through depolyploidizing divisions and regulated autophagic degradation. Cell Biol Int 2011; 35: 687−695.
Bertucci F, Finetti P, Cervera N, Esterni B, Hermitte F, Viens P, et al. How basal are triple-negative breast cancers? Int J Cancer 2008; 123: 236−240.
Sheridan C, Kishimoto H, Fuchs RK, Mehrotra S, Bhat-Nakshatri P, Turner CH, et al. CD44+/CD24− breast cancer cells exhibit enhanced invasive properties: An early step necessary for metastasis. Breast Cancer Res 2006; 8: R59.
Mani SA, Guo W, Liao M-J, Eaton EN, Ayyanan A, Zhou AY, et al. The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell 2008; 133: 704−715.
Zhang S, Mercado-Uribe I, Xing Z, Sun B, Kuang J, Liu J. Generation of cancer stem-like cells through the formation of polyploid giant cancer cells. Oncogene 2014; 33: 116−128.
Salmina K, Jankevics E, Huna A, Perminov D, Radovica I, Klymenko T, et al. Up-regulation of the embryonic self-renewal network through reversible polyploidy in irradiated p53-mutant tumor cells. Exp Cell Res 2010; 316: 2099−2112.
Ghisolfi L, Keates AC, Hu X, Lee D, Li CJ. Ionizing radiation induces stemness in cancer cells. PLoS ONE 2012; 7: e43628.
Lagadec C, Vlashi E, Della Donna L, Dekmezian C, Pajonk F. Radiation-induced reprogramming of breast cancer cells. Stem Cells 2012; 30: 833−844.
Niu N, Mercado-Uribe I, Liu J. Dedifferentiation into blastomere-like cancer stem cells via formation of polyploid giant cancer cells. Oncogene 2017; 36: 4887−4900.
Ben-Porath I, Thomson MW, Carey VJ, Ge R, Bell GW, Regev A, et al. An embryonic stem cell-like gene expression signature in poorly differentiated human tumors. Nat Genet 2008; 40: 499−507.
Pisco AO, Huang S. Non-genetic cancer cell plasticity and therapy-induced stemness in tumour relapse: "What does not kill me strengthens me". Br J Cancer 2015; 112: 1725−1732.
Mirzayans R, Andrais B, Murray D. Roles of polyploid/multinucleated giant cancer cells in metastasis and disease relapse following anticancer treatment. Cancers 2018; 10: 118.
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