Fluid shear stress induces cancer stem cell-like phenotype in MCF7 breast cancer cell line without inducing epithelial to mesenchymal transition

Triantafillu,Ursula L.; Park,Seungjo; Klaassen,Nikki L.; Raddatz,Andrew D.; Kim,Yonghyun

doi:10.3892/ijo.2017.3865

Fluid shear stress induces cancer stem cell-like phenotype in MCF7 breast cancer cell line without inducing epithelial to mesenchymal transition

Authors:
- Ursula L. Triantafillu
- Seungjo Park
- Nikki L. Klaassen
- Andrew D. Raddatz
- Yonghyun Kim
View Affiliations

Affiliations: Department of Chemical and Biological Engineering, The University of Alabama, Tuscaloosa, AL, USA, Department of Chemical Engineering, Kansas State University, Manhattan, KS, USA
Published online on: January 30, 2017 https://doi.org/10.3892/ijo.2017.3865
Pages: 993-1001

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

Metastasis is the leading cause of cancer deaths due to the spread of cancer cells through the blood vessels and the subsequent formation of secondary tumors. Metastasizing cancer cells in the human vasculature are called circulating tumor cells (CTCs) and are characterized to express the epithelial cell adhesion molecule (EpCAM). They are further known to survive physiological fluid shear stress (FSS) conditions. However, the effect of FSS on CTC molecular phenotype, such as the epithelial to mesenchymal transition (EMT) and cancer stem cell (CSC) expression, has not been extensively studied. Here, CTCs in FSS are evaluated in an in vitro model system. MCF7 and MDA-MB-231 breast cancer cell lines were grown in adherent and suspension culture media. The cell lines were tested for EMT and CSC genetic and protein markers using qRT-PCR and flow cytometry, respectively. Suspension cells showed a significantly increased EMT signature compared to adherent cells (p<0.05), suggesting that they model cells detaching from primary tumors in vivo. Upon application of FSS, MCF7 and MDA-MB-231 cells did not show a significant change in EMT expression (p>0.05), but there was a statistically significant increase of the CSC population in MCF7 suspension cultures (p<0.05). These results with MCF7 suggest that CTCs can be modeled in vitro as non-adherent cancer cells in FSS and that they show an increased CSC-like signature during circulation, providing new insights to the importance of CSC-targeting strategies when treating metastatic patients.

View Figures

View References

1	Chaffer CL and Weinberg RA: A perspective on cancer cell metastasis. Science. 331:1559–1564. 2011. View Article : Google Scholar : PubMed/NCBI
2	Dong Y, Skelley AM, Merdek KD, Sprott KM, Jiang C, Pierceall WE, Lin J, Stocum M, Carney WP and Smirnov DA: Microfluidics and circulating tumor cells. J Mol Diagn. 15:149–157. 2013. View Article : Google Scholar
3	Li P, Stratton ZS, Dao M, Ritz J and Huang TJ: Probing circulating tumor cells in microfluidics. Lab Chip. 13:602–609. 2013. View Article : Google Scholar : PubMed/NCBI
4	Friedlander TW, Premasekharan G and Paris PL: Looking back, to the future of circulating tumor cells. Pharmacol Ther. 142:271–280. 2014. View Article : Google Scholar
5	Broersen LH, van Pelt GW, Tollenaar RA and Mesker WE: Clinical application of circulating tumor cells in breast cancer. Cell Oncol (Dordr). 37:9–15. 2014. View Article : Google Scholar
6	Mitchell MJ and King MR: Computational and experimental models of cancer cell response to fluid shear stress. Front Oncol. 3:442013. View Article : Google Scholar : PubMed/NCBI
7	Barnes JM, Nauseef JT and Henry MD: Resistance to fluid shear stress is a conserved biophysical property of malignant cells. PloS One. 7:e509732012. View Article : Google Scholar : PubMed/NCBI
8	Aktas B, Tewes M, Fehm T, Hauch S, Kimmig R and Kasimir-Bauer S: Stem cell and epithelial-mesenchymal transition markers are frequently overexpressed in circulating tumor cells of metastatic breast cancer patients. Breast Cancer Res. 11:R462009. View Article : Google Scholar : PubMed/NCBI
9	Ip CK, Li SS, Tang MY, Sy SK, Ren Y, Shum HC and Wong AS: Stemness and chemoresistance in epithelial ovarian carcinoma cells under shear stress. Sci Rep. 6:267882016. View Article : Google Scholar : PubMed/NCBI
10	Nguyen LV, Vanner R, Dirks P and Eaves CJ: Cancer stem cells: An evolving concept. Nat Rev Cancer. 12:133–143. 2012.PubMed/NCBI
11	Reya T, Morrison SJ, Clarke MF and Weissman IL: Stem cells, cancer, and cancer stem cells. Nature. 414:105–111. 2001. View Article : Google Scholar : PubMed/NCBI
12	Al-Hajj M, Wicha MS, Benito-Hernandez A, Morrison SJ and Clarke MF: Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci USA. 100:3983–3988. 2003. View Article : Google Scholar : PubMed/NCBI
13	Ginestier C, Hur MH, Charafe-Jauffret E, Monville F, Dutcher J, Brown M, Jacquemier J, Viens P, Kleer CG, Liu S, et al: ALDH1 is a marker of normal and malignant human mammary stem cells and a predictor of poor clinical outcome. Cell Stem Cell. 1:555–567. 2007. View Article : Google Scholar
14	Visvader JE and Lindeman GJ: Cancer stem cells in solid tumours: Accumulating evidence and unresolved questions. Nat Rev Cancer. 8:755–768. 2008. View Article : Google Scholar : PubMed/NCBI
15	Ponti D, Costa A, Zaffaroni N, Pratesi G, Petrangolini G, Coradini D, Pilotti S, Pierotti MA and Daidone MG: Isolation and in vitro propagation of tumorigenic breast cancer cells with stem/progenitor cell properties. Cancer Res. 65:5506–5511. 2005. View Article : Google Scholar : PubMed/NCBI
16	Liao M-J, Zhang CC, Zhou B, Zimonjic DB, Mani SA, Kaba M, Gifford A, Reinhardt F, Popescu NC, Guo W, et al: Enrichment of a population of mammary gland cells that form mammospheres and have in vivo repopulating activity. Cancer Res. 67:8131–8138. 2007. View Article : Google Scholar : PubMed/NCBI
17	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−ΔΔC(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
18	Haley EM, Tilson SG, Triantafillu UL, Magrath JW and Kim Y: Acidic pH with coordinated reduction of basic fibroblast growth factor maintains the glioblastoma stem cell-like phenotype in vitro. J Biosci Bioeng. Jan 4–2017.Epub ahead of print. View Article : Google Scholar
19	Mallini P, Lennard T, Kirby J and Meeson A: Epithelial-to-mesenchymal transition: What is the impact on breast cancer stem cells and drug resistance. Cancer Treat Rev. 40:341–348. 2014. View Article : Google Scholar
20	Lamouille S, Xu J and Derynck R: Molecular mechanisms of epithelial-mesenchymal transition. Nat Rev Mol Cell Biol. 15:178–196. 2014. View Article : Google Scholar : PubMed/NCBI
21	Onder TT, Gupta PB, Mani SA, Yang J, Lander ES and Weinberg RA: Loss of E-cadherin promotes metastasis via multiple downstream transcriptional pathways. Cancer Res. 68:3645–3654. 2008. View Article : Google Scholar : PubMed/NCBI
22	van Roy F: Beyond E-cadherin: Roles of other cadherin super-family members in cancer. Nat Rev Cancer. 14:121–134. 2014. View Article : Google Scholar : PubMed/NCBI
23	Schnell U, Cirulli V and Giepmans BN: EpCAM: Structure and function in health and disease. Biochim Biophys Acta. 1828:1989–2001. 2013. View Article : Google Scholar : PubMed/NCBI
24	Yuan X, Curtin J, Xiong Y, Liu G, Waschsmann-Hogiu S, Farkas DL, Black KL and Yu JS: Isolation of cancer stem cells from adult glioblastoma multiforme. Oncogene. 23:9392–9400. 2004. View Article : Google Scholar : PubMed/NCBI
25	Bos PD, Nguyen DX and Massagué J: Modeling metastasis in the mouse. Curr Opin Pharmacol. 10:571–577. 2010. View Article : Google Scholar : PubMed/NCBI
26	Saxena M and Christofori G: Rebuilding cancer metastasis in the mouse. Mol Oncol. 7:283–296. 2013. View Article : Google Scholar : PubMed/NCBI
27	Tang KD, Holzapfel BM, Liu J, Lee TK, Ma S, Jovanovic L, An J, Russell PJ, Clements JA, Hutmacher DW, et al: Tie-2 regulates the stemness and metastatic properties of prostate cancer cells. Oncotarget. 7:2572–2584. 2016.
28	Lee HJ and Koh GY: Shear stress activates Tie2 receptor tyrosine kinase in human endothelial cells. Biochem Biophys Res Commun. 304:399–404. 2003. View Article : Google Scholar : PubMed/NCBI
29	Green DL and Karpatkin S: Effect of cancer on platelets. Coagulation in Cancer. Kwaan CH and Green D: Springer; Boston, MA: pp. 17–30. 2009, View Article : Google Scholar
30	Gay LJ and Felding-Habermann B: Contribution of platelets to tumour metastasis. Nat Rev Cancer. 11:123–134. 2011. View Article : Google Scholar : PubMed/NCBI
31	Mitchell MJ, Denais C, Chan MF, Wang Z, Lammerding J and King MR: Lamin A/C deficiency reduces circulating tumor cell resistance to fluid shear stress. Am J Physiol Cell Physiol. 309:C736–C746. 2015.PubMed/NCBI
32	Christiansen JJ and Rajasekaran AK: Reassessing epithelial to mesenchymal transition as a prerequisite for carcinoma invasion and metastasis. Cancer Res. 66:8319–8326. 2006. View Article : Google Scholar : PubMed/NCBI
33	Reymond N, d'Água BB and Ridley AJ: Crossing the endothelial barrier during metastasis. Nat Rev Cancer. 13:858–870. 2013. View Article : Google Scholar : PubMed/NCBI
34	Nieman MT, Prudoff RS, Johnson KR and Wheelock MJ: N-cadherin promotes motility in human breast cancer cells regardless of their E-cadherin expression. J Cell Biol. 147:631–644. 1999. View Article : Google Scholar : PubMed/NCBI
35	Ricardo S, Vieira AF, Gerhard R, Leitão D, Pinto R, Cameselle-Teijeiro JF, Milanezi F, Schmitt F and Paredes J: Breast cancer stem cell markers CD44, CD24 and ALDH1: Expression distribution within intrinsic molecular subtype. J Clin Pathol. 64:937–946. 2011. View Article : Google Scholar : PubMed/NCBI
36	Mani SA, Guo W, Liao M-J, Eaton EN, Ayyanan A, Zhou AY, Brooks M, Reinhard F, Zhang CC, Shipitsin M, et al: The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell. 133:704–715. 2008. View Article : Google Scholar : PubMed/NCBI
37	Liu S, Cong Y, Wang D, Sun Y, Deng L, Liu Y, Martin-Trevino R, Shang L, McDermott SP, Landis MD, et al: Breast cancer stem cells transition between epithelial and mesenchymal states reflective of their normal counterparts. Stem Cell Rep. 2:78–91. 2013. View Article : Google Scholar
38	Sanpaolo P, Barbieri V and Genovesi D: Prognostic value of breast cancer subtypes on breast cancer specific survival, distant metastases and local relapse rates in conservatively managed early stage breast cancer: A retrospective clinical study. Eur J Surg Oncol. 37:876–882. 2011. View Article : Google Scholar : PubMed/NCBI
39	Paquet ER and Hallett MT: Absolute assignment of breast cancer intrinsic molecular subtype. J Natl Cancer Inst. 107:dju3572015. View Article : Google Scholar
40	Holliday DL and Speirs V: Choosing the right cell line for breast cancer research. Breast Cancer Res. 13:2152011. View Article : Google Scholar : PubMed/NCBI