Bystricky Branislav, Cierna Zuzana, Sieberova Gabriela, Janega Pavol, Karaba Marian, Minarik Gabriel, Benca Juraj, Sedlackova Tatiana, Jurisova Silvia, Gronesova Paulina, Pindak Daniel, Macuch Jan, Mardiak Jozef, Mego Michal
2nd Department of Medical Oncology, Faculty of Medicine, Comenius University, Bratislava, Slovakia.
Oncology Department Faculty Hospital Trencin, Trencin, Slovakia.
Anticancer Res. 2017 May;37(5):2727-2734. doi: 10.21873/anticanres.11624.
BACKGROUND/AIM: Annexin A2 (ANXA2) is a phospholipid-binding protein involved in fibrinolysis, cell proliferation, migration and metastatic dissemination. Circulating tumor cells (CTCs) are cells responsible for tumor dissemination and have a prognostic value in several types of cancers including breast cancer. Previously, we found correlation between CTCs and activation of coagulation. This study aimed to correlate CTCs with ANXA2 expression on CTCs, tumor cells and tumor associated stroma in primary breast cancer (PBC) patients.
This prospective study included 101 PBC patients treated by primary surgery. CTCs were detected by quantitative real-time polymerase chain reaction (qRT-PCR) assay for the expression of epithelial (CK19) or epithelial-mesenchymal transition (EMT) genes [TWIST1, SNAI1, SNAI2, zinc finger E-box-binding homeobox 1 (ZEB1)]. ANXA2 expression on CTCs was detected by qRT-PCR, while expression of ANXA2 in tumor specimen was evaluated by immunohistochemistry and expressed by a weighted histoscore, evaluating both the percentage of positive cells and the intensity of membrane and cytoplasmic staining. Results of hormone receptors, HER2 status, B-cell lymphoma 2 (bcl-2) protein expression and protein p53 were reported as either positive or negative on histopathology report without further quantification.
CTCs were detected in 24.8% patients. Patients with epithelial CTCs had a significantly higher ANXA2 expression on CTCs than those of patients without CTCs (p=0.01). There was no association between CTCs and ANXA2 protein expression in tumor cells. However, patients, whom CTCs with EMT phenotype were detected in, had higher ANXA2 expression in tumor stroma when compared to those with absent EMT CTCs (p=0.04). Hormone-negative tumors had significantly higher ANXA2 expression in tumor cells compared to hormone-positive tumors (p=0.03). Similarly, tumors without bcl-2 protein expression had higher tumor levels of ANXA2 compared to tumor cells that were bcl-2 positive (p=0.05).
ANXA2 stromal expression might play a key role in aggressive tumor phenotype associated with increased EMT CTCs release, however, other factors beyond ANXA2 are responsible for coagulation activation mediated by CTCs in breast cancer patients.
背景/目的：膜联蛋白A2（ANXA2）是一种磷脂结合蛋白，参与纤维蛋白溶解、细胞增殖、迁移和转移扩散。循环肿瘤细胞（CTC）是导致肿瘤扩散的细胞，在包括乳腺癌在内的多种癌症中具有预后价值。此前，我们发现CTC与凝血激活之间存在相关性。本研究旨在探讨原发性乳腺癌（PBC）患者中CTC与CTC、肿瘤细胞及肿瘤相关基质上ANXA2表达之间的相关性。
本前瞻性研究纳入了101例接受初次手术治疗的PBC患者。通过定量实时聚合酶链反应（qRT-PCR）检测上皮（CK19）或上皮-间质转化（EMT）基因[TWIST1、SNAI1、SNAI2、锌指E盒结合同源框1（ZEB1）]的表达来检测CTC。通过qRT-PCR检测CTC上ANXA2的表达，而通过免疫组织化学评估肿瘤标本中ANXA2的表达，并通过加权组织学评分表示，评估阳性细胞百分比以及膜和细胞质染色强度。激素受体、HER2状态、B细胞淋巴瘤2（bcl-2）蛋白表达和p53蛋白的结果在组织病理学报告中报告为阳性或阴性，无需进一步定量。
24.8%的患者检测到CTC。上皮性CTC患者的CTC上ANXA2表达明显高于无CTC患者（p=0.01）。CTC与肿瘤细胞中ANXA2蛋白表达之间无关联。然而，检测到具有EMT表型CTC的患者，其肿瘤基质中ANXA2表达高于无EMT CTC的患者（p=0.04）。激素阴性肿瘤的肿瘤细胞中ANXA2表达明显高于激素阳性肿瘤（p=0.03）。同样，与bcl-2阳性的肿瘤细胞相比，无bcl-2蛋白表达的肿瘤中ANXA2水平更高（p=0.05）。
ANXA2基质表达可能在与EMT CTC释放增加相关的侵袭性肿瘤表型中起关键作用，然而，除ANXA2外的其他因素导致了乳腺癌患者中由CTC介导的凝血激活。
