Background: This is a feasibility study to determine whether circulating tumour cells (CTCs) are detectable and suitable for molecular profiling in advanced endometrial cancer (aEC). Method: Between October 2012 and February 2014, 30 patients with aEC had baseline and up to 3 follow-up samples. CTCs and stathmin expression were evaluated using the CellSearch platform. Epithelial cell adhesion molecule (EpCAM) and stathmin immunohistochemistry were performed on FFPE tumour tissue. Results: Eighteen from 30 (60%) patients had detectable CTCs during study [1 CTC (n = 7), 2 (n = 4), 3 (n = 1), 4 (n = 2), 7 (n = 1), 8 (n = 1), 22 (n = 1), 172 (n = 1) in 7.5 ml blood]. Ten from 18 patients had between 50 and 100% of detectable CTCs that were stathmin positive. More CTC-positive than CTC-negative patients had non-endometrioid versus endometrioid histology, tumour size ≥5 versus <5 cm, higher-stage disease and worse survival [hazard ratio 3.3, p > 0.05, 95% confidence interval 0.7-16.2]. Twenty-one tumour blocks were tested for EpCAM and stathmin immunohistochemistry (IHC). Stathmin tumour immunostaining scores (TIS) on IHC were higher in CTC-positive patients. Conclusion: CTC enumeration and molecular profiling with stathmin on the CellSearch platform is feasible in aEC. Stathmin TIS on IHC, a known prognostic marker in EC, was associated with CTC positivity.

Keywords:
Circulating tumour cells, Endometrial cancer, Stathmin, CellSearch, Epithelial cell adhesion molecule
1.
Cancer Statistics Registrations, England (Series MB1) - No 43. 2012; http://www.ons.gov.uk/ons/rel/vsob1/cancer-statistics-registrations-england-series-mb1-/index.html.
2.
Allard WJ, et al: Tumor cells circulate in the peripheral blood of all major carcinomas but not in healthy subjects or patients with nonmalignant diseases. Clin Cancer Res 2004;10:6897-6904.
3.
Attard G, et al: Characterization of ERG, AR and PTEN gene status in circulating tumor cells from patients with castration-resistant prostate cancer. Cancer Res 2009;69:2912-2918.
4.
Gasch C, et al: Heterogeneity of epidermal growth factor receptor status and mutations of KRAS/PIK3CA in circulating tumor cells of patients with colorectal cancer. Clin Chem 2013;59:252-260.
5.
Marchetti A, et al: Assessment of EGFR mutations in circulating tumor cell preparations from NSCLC patients by next generation sequencing: toward a real-time liquid biopsy for treatment. PLoS One 2014;9:e103883.
6.
Shaffer DR, et al: Circulating tumor cell analysis in patients with progressive castration-resistant prostate cancer. Clin Cancer Res 2007;13:2023-2029.
7.
de Bono JS, et al: Circulating tumor cells predict survival benefit from treatment in metastatic castration-resistant prostate cancer. Clin Cancer Res 2008;14:6302-6309.
8.
Bidard FC, et al: Clinical validity of circulating tumour cells in patients with metastatic breast cancer: a pooled analysis of individual patient data. Lancet Oncol 2014;15:406-414.
9.
Cohen SJ, et al: Relationship of circulating tumor cells to tumor response, progression-free survival, and overall survival in patients with metastatic colorectal cancer. J Clin Oncol 2008;26:3213-3221.
10.
Went P, Dirnhofer S, Schopf D, Moch H, Spizzo G: Expression and prognostic significance of EpCAM. J Cancer Mol 2008;3:6.
11.
Rubin CI, Atweh GF: The role of stathmin in the regulation of the cell cycle. J Cell Biochem 2004;93:242-250.
12.
Trovik J, et al: Stathmin overexpression identifies high-risk patients and lymph node metastasis in endometrial cancer. Clin Cancer Res 2011;17:3368-3377.
13.
Baquero MT, et al: Stathmin expression and its relationship to microtubule-associated protein tau and outcome in breast cancer. Cancer 2012;118:4660-4669.
14.
Friedrich B, et al: Differentiation-stage specific expression of oncoprotein 18 in human and rat prostatic adenocarcinoma. Prostate 1995;27:102-109.
15.
Golouh R, et al: The prognostic value of Stathmin-1, S100A2, and SYK proteins in ER-positive primary breast cancer patients treated with adjuvant tamoxifen monotherapy: an immunohistochemical study. Breast Cancer Res Treat 2008;110:317-326.
16.
Ke B, et al: Overexpression of stathmin 1 is associated with poor prognosis of patients with gastric cancer. Tumour Biol 2013;34:3137-3145.
17.
Kouzu Y, et al: Overexpression of stathmin in oral squamous-cell carcinoma: correlation with tumour progression and poor prognosis. Br J Cancer 2006;94:717-723.
18.
Lin WC, et al: Expression of stathmin in localized upper urinary tract urothelial carcinoma: correlations with prognosis. Urology 2009;74:1264-1269.
19.
Salvesen HB, et al: Integrated genomic profiling of endometrial carcinoma associates aggressive tumors with indicators of PI3 kinase activation. Proc Natl Acad Sci USA 2009;106:4834-4839.
20.
Su D, et al: Stathmin and tubulin expression and survival of ovarian cancer patients receiving platinum treatment with and without paclitaxel. Cancer 2009;115:2453-2463.
21.
Trovik J, et al: Stathmin is superior to AKT and phospho-AKT staining for the detection of phosphoinositide 3-kinase activation and aggressive endometrial cancer. Histopathology 2010;57:641-646.
22.
Xi W, et al: Expression of stathmin/op18 as a significant prognostic factor for cervical carcinoma patients. J Cancer Res Clin Oncol 2009;135:837-846.
23.
Alli E, et al: Effect of stathmin on the sensitivity to antimicrotubule drugs in human breast cancer. Cancer Res 2002;62:6864-6869.
24.
Werner HM, et al: Stathmin protein level, a potential predictive marker for taxane treatment response in endometrial cancer. PLoS One 2014;9:e90141.
25.
Werner HM, Trovik J, Wik E, Akslen LA, Salvesen HB: Stathmin expression predicts response to taxanes in metastatic endometrial cancer (abstract). Cancer Res 2011;71(suppl 1);3158.
26.
Saal LH, et al: Poor prognosis in carcinoma is associated with a gene expression signature of aberrant PTEN tumor suppressor pathway activity. Proc Natl Acad Sci USA 2007;104:7564-7569.
27.
Engelsen IB, et al: Pathologic expression of p53 or p16 in preoperative curettage specimens identifies high-risk endometrial carcinomas. Am J Obstet Gynecol 2006;195:979-986.
28.
Spizzo G, et al: EpCAM expression in primary tumour tissues and metastases: an immunohistochemical analysis. J Clin Pathol 2011;64:415-420.
29.
Salvesen HB, Das S, Akslen LA: Loss of nuclear p16 protein expression is not associated with promoter methylation but defines a subgroup of aggressive endometrial carcinomas with poor prognosis. Clin Cancer Res 2000;6:153-159.
30.
Vandenput I, et al: Evolution in endometrial cancer: evidence from an immunohistochemical study. Int J Gynecol Cancer 2011;21:316-322.
31.
Baselga J: Targeting the phosphoinositide-3 (PI3) kinase pathway in breast cancer. Oncologist 2011;16(suppl 1):12-19.
32.
Majumder PK, Sellers WR: Akt-regulated pathways in prostate cancer. Oncogene 2005;24:7465-7474.
33.
Bogani G, et al: Detection of circulating tumor cells in high-risk endometrial cancer. Anticancer Res 2015;35:683-687.
34.
Spizzo G, et al: Overexpression of epithelial cell adhesion molecule (Ep-CAM) is an independent prognostic marker for reduced survival of patients with epithelial ovarian cancer. Gynecol Oncol 2006;103:483-488.
35.
Rao CG, et al: Expression of epithelial cell adhesion molecule in carcinoma cells present in blood and primary and metastatic tumors. Int J Oncol 2005;27:49-57.
36.
Saucedo-Zeni N, et al: A novel method for the in vivo isolation of circulating tumor cells from peripheral blood of cancer patients using a functionalized and structured medical wire. Int J Oncol 2012;41:1241-1250.
37.
Vona G, et al: Isolation by size of epithelial tumor cells: a new method for the immunomorphological and molecular characterization of circulating tumor cells. Am J Pathol 2000;156:57-63.
38.
Paterlini-Brechot P, Benali NL: Circulating tumor cells (CTC) detection: clinical impact and future directions. Cancer Lett 2007;253:180-204.
39.
Gascoyne PR, et al: Isolation of rare cells from cell mixtures by dielectrophoresis. Electrophoresis 2009;30:1388-1398.
40.
Fuchs AB, et al: Electronic sorting and recovery of single live cells from microlitre sized samples. Lab Chip 2006;6:121-126.
41.
Lopez-Riquelme N, et al: Imaging cytometry for counting circulating tumor cells: comparative analysis of the CellSearch vs ImageStream systems. APMIS 2013;121:1139-1143.
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2016
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