Comprehensive Analysis of CXCR4, JUNB, and PD-L1 Expression in Circulating Tumor Cells (CTCs) from Prostate Cancer Patients
Argyro Roumeliotou,
Areti Strati,
Foteini Chamchougia,
Anastasia Xagara,
Victoria Tserpeli,
Stavroula Smilkou,
Elina Lagopodi,
Athina Christopoulou,
Emmanouil Kontopodis,
Ioannis Drositis,
Nikolaos Androulakis,
Vassilis Georgoulias,
Filippos Koinis,
Athanasios Kotsakis,
Evi Lianidou,
Galatea Kallergi
Affiliations
Argyro Roumeliotou
Laboratory of Biochemistry/Metastatic Signaling, Department of Biology, University of Patras, 26504 Patras, Greece
Areti Strati
Analysis of Circulating Tumor Cells Lab, Laboratory of Analytical Chemistry, Department of Chemistry, National and Kapodistrian University of Athens, 15771 Athens, Greece
Foteini Chamchougia
Laboratory of Biochemistry/Metastatic Signaling, Department of Biology, University of Patras, 26504 Patras, Greece
Anastasia Xagara
Faculty of Medicine, School of Health Sciences, University of Thessaly, 41500 Larissa, Greece
Victoria Tserpeli
Analysis of Circulating Tumor Cells Lab, Laboratory of Analytical Chemistry, Department of Chemistry, National and Kapodistrian University of Athens, 15771 Athens, Greece
Stavroula Smilkou
Analysis of Circulating Tumor Cells Lab, Laboratory of Analytical Chemistry, Department of Chemistry, National and Kapodistrian University of Athens, 15771 Athens, Greece
Elina Lagopodi
Analysis of Circulating Tumor Cells Lab, Laboratory of Analytical Chemistry, Department of Chemistry, National and Kapodistrian University of Athens, 15771 Athens, Greece
Athina Christopoulou
Oncology Unit, ST Andrews General Hospital of Patras, 26332 Patras, Greece
Emmanouil Kontopodis
Department of Oncology, Venizeleion General Hospital of Heraklion, 71409 Heraklion, Greece
Ioannis Drositis
Department of Oncology, Venizeleion General Hospital of Heraklion, 71409 Heraklion, Greece
Nikolaos Androulakis
Department of Oncology, Venizeleion General Hospital of Heraklion, 71409 Heraklion, Greece
Vassilis Georgoulias
Hellenic Oncology Research Group, 11526 Athens, Greece
Filippos Koinis
Faculty of Medicine, School of Health Sciences, University of Thessaly, 41500 Larissa, Greece
Athanasios Kotsakis
Faculty of Medicine, School of Health Sciences, University of Thessaly, 41500 Larissa, Greece
Evi Lianidou
Analysis of Circulating Tumor Cells Lab, Laboratory of Analytical Chemistry, Department of Chemistry, National and Kapodistrian University of Athens, 15771 Athens, Greece
Galatea Kallergi
Laboratory of Biochemistry/Metastatic Signaling, Department of Biology, University of Patras, 26504 Patras, Greece
CXCR4, JUNB and PD-L1 are implicated in cancer progression and metastasis. The current study investigated these biomarkers in CTCs isolated from metastatic prostate cancer (mPCa) patients at the RNA and protein levels. CTCs were isolated from 48 mPCa patients using the Ficoll density gradient and ISET system (17 out of 48). The (CK/PD-L1/CD45) and (CK/CXCR4/JUNB) phenotypes were identified using two triple immunofluorescence stainings followed by VyCAP platform analysis. Molecular analysis was conducted with an EpCAM-dependent method for 25/48 patients. CK-8, CK-18, CK-19, JUNB, CXCR4, PD-L1, and B2M (reference gene) were analyzed with RT-qPCR. The (CK+/PD-L1+/CD45-) and the (CK+/CXCR4+/JUNB+) were the most frequent phenotypes (61.1% and 62.5%, respectively). Furthermore, the (CK+/CXCR4+/JUNB-) phenotype was correlated with poorer progression-free survival [(PFS), HR: 2.5, p = 0.049], while the (CK+/PD-L1+/CD45-) phenotype was linked to decreased overall survival [(OS), HR: 262.7, p = 0.007]. Molecular analysis revealed that 76.0% of the samples were positive for CK-8,18, and 19, while 28.0% were positive for JUNB, 44.0% for CXCR4, and 48.0% for PD-L1. Conclusively, CXCR4, JUNB, and PD-L1 were highly expressed in CTCs from mPCa patients. The CXCR4 protein expression was associated with poorer PFS, while PD-L1 was correlated with decreased OS, providing new biomarkers with potential clinical relevance.
