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Международный эндокринологический журнал Том 22, №1, 2026

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Рівень ZEB1 у тканині папілярної карциноми щитоподібної залози, метастазах, мононуклеарних клітинах крові та плазмі

Рівень ZEB1 у тканині папілярної карциноми щитоподібної залози, метастазах, мононуклеарних клітинах крові та плазмі

Авторы: P.P. Zinych, V.M. Pushkarev, N.I. Levchuk, Ye.A. Shelkovoy, M.Yu. Bolgov
State Institution “V.P. Komisarenko Institute of Endocrinology and Metabolism of the NAMS of Ukraine”, Kyiv, Ukraine

Рубрики: Эндокринология

Разделы: Клинические исследования

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Резюме

Актуальність. Епітеліально-мезенхімальний перехід (ЕМП) є ключовим процесом у поширенні пухлинних клітин. Білок ZEB1 є центральним транскрипційним регулятором ЕМП — клітинної програми, яка лежить в основі інвазії пухлини, метастатичної дисемінації та терапевтичної резистентності. Дані свідчать про те, що аберантна активація ZEB1 обумовлює епітеліальну пластичність, інвазивну поведінку й метастатичну компетентність клітин раку щитоподібної залози. Мета: порівняти експресію ZEB1 в умовно нормальній тканині щитоподібної залози, доброякісних новоутвореннях, пухлинній тканині папілярної карциноми щитоподібної залози (ПКЩЗ) і мета­стазах, а також у клітинах і плазмі крові пацієнтів із ПКЩЗ. Матеріали та методи. Плазму крові, мононуклеарні клітини, післяопераційні зразки пухлинної тканини, метастазів та умовно нормальної тканини отримали у хірургічному відділенні клініки інституту. Концентрацію ZEB1 визначали за допомогою наборів для імуноферментного аналізу. Статистичний аналіз і візуалізацію даних здійснювали за допомогою програмного забезпечення Origin 2019b. Результати. Кількість ZEB1 у пухлинній тканині перевищує таку в умовно нормальній (гістологічно незміненій) тканині більше ніж у 5 разів. Приблизно таке співвідношення рівня ZEB1 між нормальною і пухлинною тканинами спостерігається у зразках пацієнтів із ПКЩЗ та метастазами. Концентрація транскрипційних факторів у метастазах перевищує норму майже в 10 разів. Кількість ZEB1 у тканині зоба не відрізняється від такої в умовно нормальній тканині пацієнтів із ПКЩЗ, але суттєво відрізняється від показника пухлинної тканини карциноми. Висновки. Наші дані свідчать про значні відмінності в концентрації ZEB1 між умовно нормальною та пухлинною тканинами щитоподібної залози. Рівень ZEB1 у метастазах був ще вищим. Слід підкреслити той факт, що як у клітинах крові, так і в плазмі пацієнтів із метастатичною ПКЩЗ рівень ZEB1 був значно вищим, ніж у здорових людей та осіб із ПКЩЗ без метастазів. Останнє може стати основою для перед­операційного прогнозу розвитку метастазів у пацієнтів із ПКЩЗ.

Background. Epithelial-mesenchymal transition (EMT) is a key process in the dissemination of tumor cells. Zinc finger E-box-binding homeobox 1 (ZEB1) is a central transcriptional regulator of EMT, a cellular program that underlies tumor invasion, metastatic dissemination, and therapeutic resistance. Evidence indicates that aberrant activation of ZEB1 promotes epithelial plasticity, invasive behavior, and metastatic competence of thyroid cancer cells. The purpose of the study was to compare the expression of ZEB1 in conditionally normal thyroid tissue, benign neoplasms, tumor tissue of papillary thyroid carcinoma (PTC) and metastasis, as well as in cells and blood plasma of patients with PTC. Materials and methods. Blood plasma, mononuclear cells, postoperative samples of tumor tissue, metastasis, and conditionally normal tissue were obtained from the surgical department of the Institute’s clinic. The ZEB1 concentration was determined using enzyme immunoassay kits. Statistical analysis and data presentation were performed using Origin 2019b software. Results. The amount of ZEB1 in tumor tissue exceeds its concentration in conditionally normal (histologically unchanged) tissue by more than 5 times. Approximately the same ratio in the amount of ZEB1 between normal and tumor tissues is observed in samples from patients with РТС and metastasis. The concentration of transcription factors in metastasis exceeds the norm by almost 10 times. The amount of ZEB1 in goiter tissue does not differ from conditionally normal tissue of РТС patients but it was significantly different from tumor tissue of carcinoma. Conclusions. Our data indicate significant differences in ZEB1 concentration between conditionally normal and tumor thyroid tissues. The level of ZEB1 in metastasis was even higher. It should be emphasized that both in blood cells and plasma of metastatic PТC patients, the level of ZEB1 was significantly higher than in healthy people and individuals with PТC without metastasis. The latter circumstance can become the basis for the preoperative prognosis of the development of metastasis in PTC patients.


Ключевые слова

папілярна карцинома щитоподібної залози; метастазування; білок ZEB1; епітеліально-мезенхімальний перехід

papillary thyroid carcinoma; metastasis; ZEB1; epithelial-mesenchymal transition

doi: http://dx.doi.org/10.22141/2224-0721.22.1.2026.1668

Introduction

Metastases are the leading cause of death from malignant tumors. The process of metastasis involves the exit of cancer cells from the primary tumor, their transition into the blood or other transport system and, finally, colonization and proliferation in a distant organ. Epithelial-mesenchymal transition (EMT) is a key process in the dissemination of tumor cells [1, 2].
EMT allows epithelial cells to be reorganized to become migratory cells with a mesenchymal phenotype. Disruption of intercellular junctions, loss of cell polarity, reorganization of the cytoskeleton, and increased cell motility are considered characteristic features of EMT. Therefore, in most experimental models, epithelial (E-cadherin) and mesenchymal (N-cadherin and vimentin) markers and morphological changes are considered as indicators confirming the EMT process. In malignant tumors, EMT is initiated by different signaling pathways through the regulation of transcription factors (EMT-TFs) and miRNAs [1–5].
The progression of EMT is controlled by several extracellular and intracellular signaling pathways. These signaling pathways are: matrix protein/ILK (integrin-linked kinase), Wnt/PI3K/β-catenin, TGF-β/SMAD, TGF-β/PI3K/NF-κB, TGF-β/RAS/ERK, growth factors/TAK1/MAPK, Jagged/NOTCH-ICD and Sonic Hedgehog/GLI1. EMT-TFs, including ZEB1/ZEB2 (zinc-finger E-box binding proteins), Snail, Slug (zinc finger proteins), Twist and E47 (basic helix-loop-helix transcription factors), play a key role in regulating the expression of their target genes associated with EMT and metastasis. All these processes are finely regulated by oncogenic and tumor suppressive miRNAs [2, 6].
The purpose of the study was to compare the expression of ZEB1 in conditionally normal thyroid tissue, benign neoplasms, tumor tissue (papillary thyroid carcinoma, PTC) and metastasis, as well as in blood cells and plasma.

Materials and methods

The research protocol was approved by the Bioethics Commission of the State Institution “V.P. Komisarenko Institute of Endocrinology and Metabolism of the NAMS of Ukraine”, protocol no. 26-KE dated April 10, 2019. All patients signed informed consent for the use of biomaterials for further diagnostic and scientific research.
Postoperative samples of PTC tumor tissue, metastasis, and conditionally normal (non-tumor) tissue, obtained from the surgical department of the Institute’s clinic, were used for research. Also, blood plasma and mononuclear cells (MNBC) were analyzed. Blood was obtained by standard venipuncture in EDTA tubes. Plasma was separated by centrifugation within 10 minutes after blood sampling. MNBC were collected as described earlier [7] using Histopaque 1077 (Sigma, USA). The concentration of protein in cell lysates was determined according to [8]. Blood controls were taken from healthy individuals without thyroid and comorbid diseases. Samples were stored at –80 °C until use.
The amount of ZEB1 was determined using enzyme immunoassay kits A78980 (Antibodies.com, UK). Measurements were performed at an optical wavelength of 450 nm on an immunoenzymatic plate analyzer Stat Fax 3200 (Awareness Technology, USA).
Patients with РТС, PTC + metastasis (Mts) and goiter (82 samples in total) participated in the study. Group 1 included 6 patients with PTC without metastasis, group 2 consisted of 14 patients with PTC with Mts, group 3 included 3 patients with goiter. Mean age of patients was 51.50 ± 0.93 years.
Statistical analysis and data presentation were performed using Origin 2019b software. The results of the study are presented as M ± SE. Student’s t-test was used to compare data groups. Values of p ≤ 0.05 were considered significant.

Results

Table 1 presents the results of the ZEB1 detection in the postoperative tissue of patients with РТС and nodular goiter. The amount of ZEB1 in tumor tissue exceeds that in conditionally normal (histologically unchanged) tissue by more than fivefold. A similar ratio was observed when comparing normal and tumor tissues from patients with PTC and lymph node Mts. Notably, the concentration of this transcription factor in metastatic tissue exceeded normal levels by almost tenfold. In contrast, ZEB1 expression in goiter tissue did not differ from that in conditionally normal tissue from PTC patients but was significantly lower than in carcinoma tissue.
Analysis of MNBCs from patients with PTC and PTC with Mts demonstrated no significant difference between healthy individuals and patients with PTC without metastasis (Fig. 1). However, in patients with PTC and Mts, ZEB1 levels were 2.3-fold higher than in controls and 1.9-fold higher than in PTC samples without metastases. In blood plasma from patients with PTC and Mts, ZEB1 levels were significantly elevated, exceeding those in control plasma by 2.7-fold and those in PTC samples without metastases by 1.67-fold.
Thus, significant differences were observed between PTC samples with and without metastasis.

Discussion

ZEB1 is at the forefront of TFs involved in the control of EMT. Deletion of ZEB1 in some cancer models significantly reduced the invasiveness of highly aggressive tumor cells and strongly inhibited metastasis. This suggests that deletion of Twist1 or Snai1 alone is not sufficient to inhibit EMT and that ZEB1 deletion has a much greater effect on tumor phenotype and Mts formation [9]. The ZEB family is more long-lived compared to other EMT-TFs [10]. ZEB1 is the main EMT-TF of the ZEB family and is associated with cellular plasticity, cell stemness, evasion of the immune response, resistance to treatment, dissemination and switching from a dormant to proliferative phenotype in distant metastatic sites and is a determinant of worse clinical prognosis for most types of human cancer [9, 11–13]. Beyond its role in EMT, ZEB1 promotes the acquisition of cancer stem cell-like properties, including enhanced self-renewal, tumor-initiating capacity, and resistance to therapy. In thyroid cancer, ZEB1 expression correlates with stemness markers such as SOX2 and OCT4, supporting its role in metastatic colonization and disease recurrence [14].
Recent studies consistently demonstrate elevated ZEB1 expression in advanced thyroid cancers compared with normal thyroid tissue or differentiated tumors. Analyses show that ZEB1 overexpression correlates with loss of E-cadherin, increased vimentin expression, and enhanced invasive capacity of thyroid cancer cells [15]. In papillary thyroid carcinoma, ZEB1 expression has been associated with extrathyroidal extension, lymph node metastasis, and higher tumor stage [16]. Metastatic thyroid cancer lesions often exhibit mesenchymal features and elevated ZEB1 expression, suggesting that ZEB1-mediated plasticity facilitates both dissemination and adaptation to distant microenvironments [14]. Functional studies using thyroid cancer cell lines indicate that ZEB1 knockdown restores epithelial characteristics, reduces migratory and invasive behavior, and suppresses EMT-related gene expression [16].
Thus, ZEB1 is a pivotal regulator of EMT, invasion, and metastasis in thyroid cancer. A deeper understanding of ZEB1-mediated mechanisms may facilitate the development of novel prognostic tools and targeted therapies for aggressive thyroid cancers.
One of the major challenges in endocrine surgery is the identification of reliable markers of metastasis in the management of PTC. Of particular clinical importance are markers that can be assessed at the preoperative stage. According to our data, proliferating cell nuclear antigen [17], expression of a rare isoform of ribosomal S6 kinase (p60S6K) [18], the tight junction protein ZO-1 [19], and matrix metalloproteinase-2 [20] are markers associated with increased aggressiveness and metastatic potential of thyroid tumors. In addition, multiple studies have demonstrated that activation of EMT and increased ZEB1 expression are closely associated with PTC progression and invasive behavior [11, 12, 21–26]. Thus, transcription factor ZEB1 represents a promising candidate among such markers.

Conclusions

Our data indicate pronounced differences (greater than fivefold) in ZEB1 concentration between conditionally normal thyroid tissue and tumor tissue. ZEB1 levels were even higher in metastatic lesions. Importantly, both blood cells and plasma from patients with metastatic PTC showed significantly elevated ZEB1 levels compared with healthy individuals and patients with non-metastatic PTC. This finding suggests that ZEB1 may serve as a promising preoperative prognostic marker for the development of metastases in PTC patients. Moreover, determination of ZEB1 expression may facilitate the differentiation of benign thyroid neoplasms from papillary thyroid carcinoma.
 
Received 22.10.2025
Revised 19.12.2025
Accepted 30.12.2025

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