Circulating tumor cells from patients with advanced prostate and breast cancer display both epithelial and mesenchymal markers - PubMed (original) (raw)

Circulating tumor cells from patients with advanced prostate and breast cancer display both epithelial and mesenchymal markers

Andrew J Armstrong et al. Mol Cancer Res. 2011 Aug.

Abstract

During cancer progression, malignant cells undergo epithelial-mesenchymal transitions (EMT) and mesenchymal-epithelial transitions (MET) as part of a broad invasion and metastasis program. We previously observed MET events among lung metastases in a preclinical model of prostate adenocarcinoma that suggested a relationship between epithelial plasticity and metastatic spread. We thus sought to translate these findings into clinical evidence by examining the existence of EMT in circulating tumor cells (CTC) from patients with progressive metastatic solid tumors, with a focus on men with castration-resistant prostate cancer (CRPC) and women with metastatic breast cancer. We showed that the majority (> 80%) of these CTCs in patients with metastatic CRPC coexpress epithelial proteins such as epithelial cell adhesion molecule (EpCAM), cytokeratins (CK), and E-cadherin, with mesenchymal proteins including vimentin, N-cadherin and O-cadherin, and the stem cell marker CD133. Equally, we found that more than 75% of CTCs from women with metastatic breast cancer coexpress CK, vimentin, and N-cadherin. The existence and high frequency of these CTCs coexpressing epithelial, mesenchymal, and stem cell markers in patients with progressive metastases has important implications for the application and interpretation of approved methods to detect CTCs.

PubMed Disclaimer

Conflict of interest statement

Disclosure of Potential Conflicts of Interest: Drs. Armstrong, Oltean, George, and Garcia-Blanco are listed as inventors in a related patent application (Application Number PCT/US10/50233) filed on September 24, 2010.

Figures

Figure 1

Figure 1. Co-expression of epithelial and mesenchymal proteins in CTCs from men with metastatic castration resistant prostate cancer (mCRPC)

All panels represent merged images derived from phase/DAPI, CD45/DAPI, CK/DAPI, and either vimentin(Vim)/DAPI, N-cadherin(N-cad)/DAPI expression, or O-cadherin(O-cad)/DAPI as indicated. Shown are examples of (A) a leukocyte with CD45 expression, (B) a CTC with no vimentin expression, (C) a CTC with vimentin expression, (D) a CTC with N-cadherin expression, (E) three CTCs, two with N-cadherin expression (arrowheads), (F) a CTC with O-cadherin expression and a nearby leukocyte, and (G) an additional CTC with O-cadherin expression. Scale bars represent 20 μm and were added from an image taken at identical magnification and resolution. Control cells were assayed in parallel at the same time of CTC collection and analysis with each set of patient samples and are shown in Supplementary Fig. 1.

Figure 2

Figure 2. Expression of vimentin and cytokeratin in prostate cancer metastases

Images are taken from a CT-guided targeted bone metastasis biopsy at the same time as circulating tumor cells were collected and evaluated for vimentin co-expression by immunofluorescence as described. Images are from patient 6 (top) and patient 7 (bottom), with CK (left) and vimentin (right) expression assayed by immunohistochemistry (20X magnification). Scale bars in the CK panels represent 50 μm and were added from an image taken at identical magnification and resolution.

Figure 3

Figure 3. E-cadherin and N-cadherin co-expression among CD45 negative nucleated cells from men with metastatic CRPC

All panels represent merged images derived from phase/DAPI, CD45/DAPI, E-cadherin(E-cad)/DAPI, and N-cadherin(N-cad)/DAPI expression as indicated. (A) a leukocyte with CD45 expression, (B) a CD45 negative nucleated cell, with E-cadherin and N-cadherin co-expression, (C) a CD45 negative nucleated cell with N-cadherin expression and no E-cadherin expression, (D) a CD45 negative nucleated cell with E-cadherin expression and no N-cadherin expression, and (E) a nucleated cell lacking expression of CD45, E-cadherin, and N-cadherin. Scale bars represent 20 μm and were added from an image taken at identical magnification and resolution. Control cells were assayed in parallel at the same time of CTC collection and analysis with each set of patient samples and are shown in Supplementary Figure 1.

Figure 4

Figure 4. Expression of a stem-like cell marker CD133 in CTCs from men with mCRPC

All panels represent merged images derived from phase/DAPI, CD45/DAPI, CK/DAPI, and CD133/DAPI expression as indicated. Shown are examples of (A) a leukocyte with CD45 expression, (B) a CD133 negative CTC, (C) a CD133 positive CTC, and (D) an additional example of a CD133 positive CTC. Scale bars represent 20 μm and were added from an image taken at identical magnification and resolution. Control cells were assayed in parallel at the same time of CTC collection and analysis with each set of patient samples and are shown in Supplementary Figure 1.

Similar articles

Cited by

References

    1. Hay ED, Zuk A. Transformations between epithelium and mesenchyme: normal, pathological, and experimentally induced. Am J Kidney Dis. 1995;26:678–90. - PubMed
    1. Zeisberg M, Neilson EG. Biomarkers for epithelial-mesenchymal transitions. J Clin Invest. 2009;119:1429–37. - PMC - PubMed
    1. Acloque H, Adams MS, Fishwick K, Bronner-Fraser M, Nieto MA. Epithelial-mesenchymal transitions: the importance of changing cell state in development and disease. J Clin Invest. 2009;119:1438–49. - PMC - PubMed
    1. Kalluri R, Weinberg RA. The basics of epithelial-mesenchymal transition. J Clin Invest. 2009;119:1420–8. - PMC - PubMed
    1. Blick T, Widodo E, Hugo H, et al. Epithelial mesenchymal transition traits in human breast cancer cell lines. Clin Exp Metastasis. 2008;25:629–42. - PubMed

Publication types

MeSH terms

Substances

Grants and funding

LinkOut - more resources