TY - JOUR
T1 - Molecular profiling of circulating tumor cells links plasticity to the metastatic process in endometrial cancer
AU - Alonso-Alconada, Lorena
AU - Muinelo-Romay, Laura
AU - Madissoo, Kadri
AU - Diaz-Lopez, Antonio
AU - Krakstad, Camilla
AU - Trovik, Jone
AU - Wik, Elisabeth
AU - Hapangama, Dharani
AU - Coenegrachts, Lieve
AU - Cano, Amparo
AU - Gil-Moreno, Antonio
AU - Chiva, Luis
AU - Cueva, Juan
AU - Vieito, Maria
AU - Ortega, Eugenia
AU - Mariscal, Javier
AU - Colas, Eva
AU - Castellvi, Josep
AU - Cusido, Maite
AU - Dolcet, Xavier
AU - Nijman, Hans W.
AU - Bosse, Tjalling
AU - Green, John A.
AU - Romano, Andrea
AU - Reventos, Jaume
AU - Lopez-Lopez, Rafael
AU - Salvesen, Helga B.
AU - Amant, Frederic
AU - Matias-Guiu, Xavier
AU - Moreno-Bueno, Gema
AU - Abal, Miguel
PY - 2014/9/27
Y1 - 2014/9/27
N2 - About 20% of patients diagnosed with endometrial cancer (EC) are considered high-risk with unfavorable prognosis. In the framework of the European Network for Individualized Treatment in EC (ENITEC), we investigated the presence and phenotypic features of Circulating Tumor Cells (CTC) in high-risk EC patients. Methods: CTC isolation was carried out in peripheral blood samples from 34 patients, ranging from Grade 3 Stage IB to Stage IV carcinomas and recurrences, and 27 healthy controls using two methodologies. Samples were subjected to EpCAM-based immunoisolation using the CELLection (TM) Epithelial Enrich kit (Invitrogen, Dynal) followed by RTqPCR analysis. The phenotypic determinants of endometrial CTC in terms of pathogenesis, hormone receptor pathways, stem cell markers and epithelial to mesenchymal transition (EMT) drivers were asked. Kruskal-Wallis analysis followed by Dunn's post-test was used for comparisons between groups. Statistical significance was set at p <0.05. Results: EpCAM-based immunoisolation positively detected CTC in high-risk endometrial cancer patients. CTC characterization indicated a remarkable plasticity phenotype defined by the expression of the EMT markers ETV5, NOTCH1, SNAI1, TGFB1, ZEB1 and ZEB2. In addition, the expression of ALDH and CD44 pointed to an association with stemness, while the expression of CTNNB1, STS, GDF15, RELA, RUNX1, BRAF and PIK3CA suggested potential therapeutic targets. We further recapitulated the EMT phenotype found in endometrial CTC through the up-regulation of ETV5 in an EC cell line, and validated in an animal model of systemic dissemination the propensity of these CTC in the accomplishment of metastasis. Conclusions: Our results associate the presence of CTC with high-risk EC. Gene-expression profiling characterized a CTC-plasticity phenotype with stemness and EMT features. We finally recapitulated this CTC-phenotype by over-expressing ETV5 in the EC cell line Hec1A and demonstrated an advantage in the promotion of metastasis in an in vivo mouse model of CTC dissemination and homing.
AB - About 20% of patients diagnosed with endometrial cancer (EC) are considered high-risk with unfavorable prognosis. In the framework of the European Network for Individualized Treatment in EC (ENITEC), we investigated the presence and phenotypic features of Circulating Tumor Cells (CTC) in high-risk EC patients. Methods: CTC isolation was carried out in peripheral blood samples from 34 patients, ranging from Grade 3 Stage IB to Stage IV carcinomas and recurrences, and 27 healthy controls using two methodologies. Samples were subjected to EpCAM-based immunoisolation using the CELLection (TM) Epithelial Enrich kit (Invitrogen, Dynal) followed by RTqPCR analysis. The phenotypic determinants of endometrial CTC in terms of pathogenesis, hormone receptor pathways, stem cell markers and epithelial to mesenchymal transition (EMT) drivers were asked. Kruskal-Wallis analysis followed by Dunn's post-test was used for comparisons between groups. Statistical significance was set at p <0.05. Results: EpCAM-based immunoisolation positively detected CTC in high-risk endometrial cancer patients. CTC characterization indicated a remarkable plasticity phenotype defined by the expression of the EMT markers ETV5, NOTCH1, SNAI1, TGFB1, ZEB1 and ZEB2. In addition, the expression of ALDH and CD44 pointed to an association with stemness, while the expression of CTNNB1, STS, GDF15, RELA, RUNX1, BRAF and PIK3CA suggested potential therapeutic targets. We further recapitulated the EMT phenotype found in endometrial CTC through the up-regulation of ETV5 in an EC cell line, and validated in an animal model of systemic dissemination the propensity of these CTC in the accomplishment of metastasis. Conclusions: Our results associate the presence of CTC with high-risk EC. Gene-expression profiling characterized a CTC-plasticity phenotype with stemness and EMT features. We finally recapitulated this CTC-phenotype by over-expressing ETV5 in the EC cell line Hec1A and demonstrated an advantage in the promotion of metastasis in an in vivo mouse model of CTC dissemination and homing.
KW - High-risk endometrial carcinomas
KW - Circulating tumor cells
KW - Epithelial to mesenchymal transition
KW - Stem cell
KW - ETV5
U2 - 10.1186/1476-4598-13-223
DO - 10.1186/1476-4598-13-223
M3 - Article
C2 - 25261936
SN - 1476-4598
VL - 13
JO - Molecular Cancer
JF - Molecular Cancer
M1 - 223
ER -