Background. The calpain proteolytic system plays an important role in the development of cancer. Detection of early cancer in the upper respiratory tract is often challenging, as symptoms are largely non-specific, and most cases are diagnosed at an advanced stage. Aim . To identify candidate markers of transition from premalignant lesions to invasive carcinoma, we studied mRNA expression levels of CAPN1 and CAPN2 and the total activity of calpains in the tumor tissues of patients with head and neck squamous cell carcinoma (HNSCC) and in the epithelial dysplasia-affected tissues of patients with chronic diseases of the upper respiratory tract. Materials and methods. The study included 32 patients with HNSCC (Т1-3N0-1М0) and 12 patients with chronic diseases of the upper respiratory system associated with epithelial dysplasia. The expression levels of CAPN1 and CAPN2 were assessed using real-time polymerase chain reaction (PCR). The calpain activity was determined by hydrolysis of the fluorogenic Suc-LLVY-AMC oligopeptide. Results. The mRNA expression levels of CAPN1 and CAPN2 were respectively 3 and 4 times higher in the tumor tissue of patients with HNSCC than in the tissue of patients with endothelial dysplasia in the upper respiratory tract. The level of calpain activity was 4.4 times higher in patients with HNSCC than in patients with epithelial dysplasia of different severity. Conclusion. The elevated mRNA expression levels of CAPN1 and CAPN2 and their activity in the tumor tissues of patients with HNSCC compared to patients with chronic respiratory diseases associated with epithelial dysplasia are likely to characterize a high potential for transition from precancerous lesion to cancer. To clarify the role of calpains in the carcinogenesis of HNSCC, further studies of intact tissues using animal models are required.

We analyzed the expression of genes encoding proteins involved in cytoskeleton remodeling (RND3, SNAI1, vimentin, cofilin, adenylate cyclase-associated protein 1, ezrin, and profilin) depending on the level of expression of protein phosphatase 1B (PPM1B) mRNA on the example of squamous cell carcinoma of the larynx and hypopharynx. Against the background of a high level of PPM1B expression, a significantly high level of profilin expression was noted. Metastasis correlated with the level of snai1 expression, while relapse after combination treatment was negatively associated with the level of vimentin expression. The obtained new data can reflect molecular peculiarities of the tumor growth in squamous cell carcinoma of the larynx and hypopharynx.

We analyzed the content of β-catenin fractions and activity and content of proteasomes in the tissues of patients with non-small cells lung cancer. The content of β-catenin fractions was elevated and proteasome-dependent proteolysis in the tumor tissue was enhanced in comparison with the corresponding unchanged lung tissue. A negative regression relationship of caspase-like activity of proteasomes and a positive correlation between the content of proteasomes and both fractions of β-catenin were found. We hypothesize that proteasomes are involved in the degradation of β-catenin due to caspase-like activity.

High aggressiveness of laryngeal and hypopharyngeal squamous cell carcinomas dictates the necessity of studying the molecular mechanisms of metastasis. Cancer metastasis is associated with remodeling of the cytoskeleton, which is carried out by actin-binding proteins (ABPs). Currently, there is insufficient data on the feasibility of determining the level of circulating ASBs in lymphogenous metastasis of laryngeal cancer (LC). Material and Methods . Blood serum analysis was carried out in 42 LC patients and 15 healthy volunteers using ELISA kits on a microplate ELISA reader Multiskan FC 100 (ThermoFisher Scientific). Among ASBs, cofilin1 (CFL1), fascin1 (FSCN1), ezrin (EZR), profilin1 (PFN1), adenylyl cyclase associated protein 1 (CAP1) were studied. Statistical processing of the results was performed using the Statistica 6.0 software package. Results . It was shown that the serum level of CAP1 was significantly higher in patients with LC compared with the group of healthy volunteers (p=0.00). As the size of the primary tumor increased, the levels of FSCN1, CAP1 and PFN1 significantly increased. The level of FSCN1 was 10 times higher and the level of CAP1 was 40% higher in LC patients with metastases than in LC patients without metastases. Thus, among the studied ASBs, FSCN1, CAP1, and PFN1 play an important role in the pathogenesis of LC. Conclusion . The results of the serum level of ASB in LC were obtained for the first time, and they determine the fundamental basis for the development of new methods for predicting the development of metastases.

We analyzed the association of the level of mRNA expression of the main endocytosis receptor LRP1 and actin-binding proteins (ezrin, profilin-1, cofilin-1, and adenylyl cyclase-associated protein 1) with the development and metastasis of laryngeal and laryngopharyngeal squamous cell carcinoma. The mRNA expression was evaluated in paired tissue samples using quantitative reverse transcription real-time PCR (RT-qPCR) and SYBR Green reagents. The study included 38 patients with stage T1-4N0-1M0 laryngeal and laryngopharyngeal squamous cell carcinoma and 10 patients with chronic hyperplastic laryngitis or grade II-III epithelial dysplasia. The expression of LRP1 in patients with laryngeal and laryngopharyngeal squamous cell carcinoma depended on the stage of the tumor process. Against the background of low expression of LRP1 mRNA, the relationship between cofilin 1 and profilin 1 expression became stronger ( r =0.08; p =0.05) and a correlation between cofilin 1 and esrin expression ( r =0.7; p =0.05) appeared. Studies on a larger patient cohort are required to make a definite conclusion on the role of LRP1 in the development of laryngeal and laryngopharyngeal squamous cell carcinoma.

Metabolic reprogramming is one of the central features of transformed cells. Elucidation of interactions between oncogenic signaling and cell metabolic processes has become the basis for extensive studies of metabolism reprogramming in tumor tissue. The review summarizes the key results of studies on the catabolic and anabolic rearrangements in tumor cells with special emphasis on carbohydrate, lipid, amino acid, and acetate metabolism determining the cancer phenotype of cells.

The aim of this study was to compare the activity of proteasomes with different variants of tumor progression in luminal and triple-negative breast cancer (BC). Materials and methods. 132 patients with primary luminal and triple-negative breast cancer in stage Т 1-3 N0 2 M 0 who had not received neoadjuvant treatment were included in this study. Proteasome chymotrypsin-like (ChTL) and caspase-like (CL) activities were determined by hydrolysis of fluorogenic peptides Suc-LLVY-AMC and Z-LLGlu-AMC in samples of tumor and adjacent tissues. The coefficients of chymotrypsin-like (kChTL) and caspase-like (kCL) proteasome activity were also calculated as the ratio of the corresponding activity in the tumor tissue to activity in the adjacent tissue. Results: Results. The increased level of ChTL and CL proteasomal activity in the tumor were observed in comparison with adjacent tissues for all molecular subtypes of BC. The process of lymphogenic metastasis of luminal A and luminal B subtypes of breast cancer is associated with significant changes in the CL activity of the proteasome. Caspase-like activity of proteasomes was increased in luminal A breast cancer with extensive lymphogenic metastasis (N2), and on the other hand it was decreased in the luminal B subtype of cancer. For the hematogenic pathway of breast cancer progression the ratio of proteasomal activity in the tumor and adjacent tissues plays a significant role. The increase in proteasome activity in the tumor over the activity of proteasomes in adjacent tissues more than 2 times was associated with poor metastatic-free prognosis of the disease.

The model of head and neck squamous cell carcinoma (HNSCC) was used to study the expression of genes encoding actin-binding proteins depending on the type of cell motility. The expression of SNAIL1 and CAPN2 mRNA in HNSCC tissue was higher than in specimens of dysplastic epithelium of the larynx and hypopharynx, which can be explained by activation of mesenchymal and amoeboid types of cell motility. In biopsy material of HNSCC patients with T1-2N0M0, expression of genes responsible for actin-binding proteins differed from that of patients with pretumor pathology of the larynx and hypopharynx: expression of FSCN was lower, while expressions of EZR and CAP1 were higher. The data attest that progression of HNSCC is associated with activation of both types of cell motility and with the changes in the expression of mRNA encoding cell motility proteins.

Chimotrypsin-like (ChTL) and caspase-like (CL) proteasomal activities were investigated in different variants of the tumor progression of luminal breast cancer. Patients with primary luminal breast cancer (n = 123) in stage T1-3N0-2M0 who had not received neoadjuvant treatment were included in this study. Proteasome ChTL and CL activities were determined in the samples of tumor and adjacent tissues. The coefficients of chymotrypsin-like (kChTL) and caspase-like (kCL) proteasome activity were also calculated as the ratio of the corresponding activity in the tumor tissue to activity in the adjacent tissue. ChTL, CL, kChTL and kCL in the tissues of luminal A and B breast cancer with lymphogenic metastasis were compared, and their association with hematogenous metastasis was evaluated. On the one hand, CL activity of proteasomes increased in luminal A breast cancer with extensive lymphogenic metastasis (N2), on the other hand it decreased in the luminal B subtype of cancer. The ratio of proteasomal activity in the tumor and adjacent tissues plays a significant role in the hematogenic pathway of breast cancer progression and is associated with poor metastatic-free survival.

The characteristics of calpain and proteasome systems in different molecular subtypes of breast cancer were studied. Tumor samples were obtained from 147 breast cancer patients, who were not previously treated with neoadjuvant therapy. Changes in the chymotrypsin-like proteasome activity and the correlation between proteasome and calpain activities were shown in different molecular subtypes of breast cancer. No changes in the calpain activity and proteasome subunits specific for various molecular subtypes of breast cancer were found. Our results show that the proteasome function undergoes changes depending on the receptor expression by tumor cells and this aspect should be further investigated.

Proteasome ubiquitin system is the important system of intracellular proteolysis. The activity of the proteasomes may undergo changes during cancer development. We studied the chymotrypsin-like activity of proteasomes, their subunit composition, and their association with tumor stage in breast cancer, head and neck squamous cell carcinoma, endometrial cancer, renal cancer, bladder cancer, stomach cancer, ovarian cancer, and colorectal cancer. The increase in chymotrypsin-like activity of proteasomes and decrease in total proteasome pool compared with adjacent tissues were shown in all malignant tumors excluding kidney cancer. The increase in chymotrypsin-like activity of proteasomes was found in primary tumors with all types of metastasis: lymphogenous of head and neck squamous cell carcinoma, intraperitoneal metastasis of ovarian cancer, hematogenous metastasis colorectal cancer. The exception was kidney cancer, in which there was a decrease in chymotrypsin-like activity with distant metastasis.

In patients with breast cancer and lung cancer, chymotrypsin-like and caspase-like activities of proteasomes and total activity of calpains in the primary tumor nodes and lymphogenic metastasis are elevated in comparison with the corresponding normal tissues. The development of lymphogenic metastases of breast cancer and lung cancer was associated with opposite change in caspase-like activity of proteasomes. These results can be useful for the development of methods for evaluation of aggressiveness of breast and lung cancer.

Резюме: известно, что злокачественный фенотип опухоли формируется в результате изменения содержания патогенетически значимых белков в клетках, на которое оказывают влияние процессы протеиновой деградации. Поэтому изучение протеолиза и выявление компонентов протеолитических систем, ассоциированных с онкологическим процессом, перспективно для поиска маркеров злокачественной трансформации и опухолевой прогрессии. В обзоре проанализированы новые возможности определения активности и содержания компонентов внутриклеточных протеолитических систем – протеасом и кальпаинов для формирования групп онкологического риска среди больных гиперплазией эндометрия, прогноза метастазирования при плоскоклеточном раке головы и шеи, раке почки, раке яичников, а также предсказания эффективности противоопухолевой терапии. Использование этих новых маркеров в онкологической практике может расширить возможности для персонализированного подхода к лечению больных.

This review presents the structure of the proteasome system, discusses its involvement in the pathogenesis of many cancers, including the process of metastasis. It describes the distinctive structural characteristics of intra- and extracellular pools of proteasomes, possible ways of their exit into the extracellular space and their forms of existence outside the cell. Also in the present review discusses the importance of circulating proteasomes for prognosis course of the cancer.

The paper discusses the capability for active movement in an extracellular matrix, wherein remodeling of the cytoskeleton by actin binding proteins plays a significant role in metastases formation. We studied the expression of actin binding proteins and β-catenin in tissues of primary tumors and metastases of ovarian and breast cancer. Contents of p45 Ser β-catenin and the actin severing protein gelsolin were decreased in metastases of ovarian cancer relative to primary tumors. The level of the cofilin, functionally similar to gelsolin, was significantly higher in metastases compared to primary ovarian and breast tumor tissue. In breast cancer, significant increase in the number of an actin monomer binder protein thymosin-β4 was observed in metastases as compared to primary tumors. The data obtained suggest the involvement of locomotor proteins in metastases formation in ovarian and breast cancer.

The chymotrypsin-like and caspase-like proteasome activities were studied in tumor tissues of 74 patients with breast cancer (BC) NМin association with tumor size and lymph node involvement. Chymotrypsin-like and caspase-like proteasome activities were Т1-40-30 higher in breast cancer tissue than in adjacent tissues. We showed that advanced BC is accompanied by increase in chymotrypsin-like and caspase-like proteasome activities. In tumors with lymph node dissemination, proteasome activity was significantly decreased, which may have prospective prognostic value. Taken together, our data indicate an important role of intracellular proteolysis in BC progression.

This review presents the structure of the proteasome system, discusses its involvement in the pathogenesis of many cancers, including the process of metastasis. It describes the distinctive structural characteristics of intra- and extracellular pools of proteasomes, possible ways of their exit into the extracellular space and their forms of existence outside the cell. Also in the present review discusses the importance of circulating proteasomes for prognosis course of the cancer.