Experimental oncology最新文献

All cancers are diseases of the genome, since the cancer cell genome typically consists of 10,000s of passenger alterations, 5-10 biologically relevant alterations, and 1-2 "actionable" alterations. Therefore, somatic mutations in cancer cells can have diagnostic, prognostic, and predictive value. Traditional methods are widely used for testing, such as immunohistochemistry, Sanger sequencing, and allele-specific PCR. However, due to the low throughput, these methods are focused exclusively on testing the most common mutations in target genes. The modern next generation sequencing (NGS) is a technology that enables precision oncology in its current form. ESCAT and ESMO Guidelines defined NGS for routine use in patients with advanced cancers such as non-squamous non-small cell lung cancer, prostate cancer, ovarian cancer, and cholangiocarcinoma. The high sensitivity of the NGS method allows it to be used to search for specific mutations in circulating tumor DNA in blood plasma and other body fluids. NGS testing has evolved from hotspot panels, actionable gene panels, and disease-specific panels to more comprehensive panels. The exome and whole genome sequencing approaches are just beginning to emerge, that is why panel-based testing remains most optimal in oncology practice. NGS is also widely used to identify new and rare mutations in cancer genes and detect inherited cancer mutations.

Background: It has been proven that changes in the morphology, representation, and organization of collagen fibers contribute to the formation of a unique microenvironment, which is associated with the metastatic potential of malignant neoplasms due to the initiation of cell migration and changes in polarization. Among the modulators of the collagen stroma, fibroblasts remain the most widely studied today. At the same time, much less attention is focused on the study of immune cells in the tumor microenvironment, in particular, mast cells (MCs).

Aim: To investigate the relationship between the MCs status and the features of the collagen matrix of breast cancer (BCa).

Materials and methods: The study was conducted on the postoperative material of 78 patients with BCa stage I-II. MCs were assessed by a histochemical method using toluidine blue. For estimation of the functional activity of MCs, a degranulation index was calculated. COL1A1, COL3A1, and MMP-9 expression in tumor tissue was assessed immunohistochemically. A visualization of collagen fibers was performed using the staining by Malory. Microphotographs were pre-processed in Adobe Photoshop SS 2019 and analyzed using the software packages CurveAlign v. 4.0 and ImageJ.

Results: Tumor tissue with a high density and functional activity of MCs was characterized by an increased expression of COL1A1 (p < 0.05), COL3A1 (p < 0.05), and MMP-9 (p < 0.05). In BCa tissue with the lower MCs degranulation index, collagen fibers become thicker (p < 0.05), shorter (p < 0.05), and denser (p < 0.05). At the same time, the existence of a relationship between the levels of miR-155-5p and the expression of COL1A1 (r = 0.703, p = 0.009), COL3A1 (r = 0.603, p = 0.043), and MMP-9 in tumor cells (r = 0.562, p = 0.039) and in the stroma (r = 0.546, p = 0.038), as well as the associations of the levels of this miRNA with the fiber length (r = -0.632, p = 0.013), width (r = -0.522, p = 0.048), and density (r = 0.699, p = 0.014) were found. Significantly higher rates of miR-155-5p expression (p < 0.05) were recorded in BCa tissue with a high index of MCs degranulation.

Conclusion: During the BCa progression, the role of MCs in the manifestation of the tumor development increases. A growing number of infiltrated MCs contributes to the activation of MMP and fibrillar collagen expression. These changes lead to increased remodeling of the tumor stroma, which is directly reflected in the spatial organization of the collagen matrix. The increased activity of proteases causes a decrease in the length and width of fibrils, which is explained by a decrease in the number of mature fibers and their disorganization in three-dimensional space. The obtained data allow us to assert that MCs play a key role not only in the formation of a specific immune microenvironment of BCa but also in determining the direction of change

Background: The quality of life (QoL) of the population determines the economic and social (including demographic) behavior of the population. It is a complex concept that has its objective and subjective dimensions. Components of the objective measurement are the individual income of the population and the income of the state. The level of the development of an individual can be determined by the level of education (or the number of years of education). The overall indicator of the country's development is the life expectancy of its citizens.

Aim: To investigate at the state level the interrelationship and interdependence of such characteristics of the QoL as the level of income, education, and life expectancy. Information Base and Methods. As an information base, we used the data from international organizations (the United Nations, the World Bank, the World Health Organization, the State Statistics Service of Ukraine) and the specialized database on mortality (Human Mortality Database) and applied general scientific and statistical research methods.

Results: The relationship between life expectancy at birth and the duration of education and domestic national income was established based on the analysis of the data of 2022 for 193 countries. The conditionality of cancer mortality from the QoL was identified. Three groups of European countries different in terms of both the QoL and the level of cancer mortality were distinguished. In these groups, the specifics of the distribution of cancer-related mortality by gender and age was analyzed.

Conclusions: The results of the study evidence that the mortality rate of the population depends on the quality of its life, in particular, well-being and education. The increase in well-being is reflected in the increase in the specific part of those who died from cancer. However, these deaths will peak later, ultimately contributing to population growth.

Background: Cervical cancer is a major health concern, with human papillomaviruses (HPV) infection being a key risk factor. However, not all HPV-infected individuals develop cancer, suggesting the additional factors may be involved. This study aims to evaluate the differences in the miR-155 and -205 expression in cervical tissue with dysplasia depending on the presence of HPV and confirmed cancer diagnosis.

Materials and methods: The expression of miR-155 and -205 in 30 formalin-fixed paraffin-embedded primary cervical tissue biopsy samples was evaluated using RT-PCR.

Results: The expression levels of miRNA-155 and -205 in cervical dysplasia samples without malignant transformation was lower than these in carcinoma in situ tissues (0.74 ± 0.21 and 1.65 ± 0.42 vs. 1.37 ± 0.18 and 2.35 ± 0.32, respectively). In carcinoma in situ cases, we found higher levels of miRNA-155 and -205 (1.6 and 1.38 times, respectively) in CIN-3/ HSIL samples compared to CIN-2/HSIL samples. The expression of both miRNAs tended to increase in HPV-positive cases and in the presence of malignant transformation compared to HPV-negative dysplasia and dysplasia without signs of malignant transformation, respectively.

Conclusions: The obtained data indicate a potential relationship between the presence of HPV infection and the expression profile of miRNA-155 and -205.

Background: This study is based on the idea of using tumor cell membrane lysis induced by diarylethene-containing analog of cytotoxic peptides (CPs) - gramicidin S to create a new approach for obtaining dendritic cells (DCs)-based anticancer vaccine. It is supposed that cancer cells undergoing immunogenic cell death release the damage-associated molecular patterns (DAMPs), and thus enhance immunogenic maturation and activation of DCs. The aim of this study is to analyze the phenotypic and functional characteristics of the generated monocyte-derived DCs loaded with CPs-treated lysates of tumor cells.

Materials and methods: The triple-negative human breast cancer cell line MDA-MB-231 was used in the study. DCs were generated from peripheral blood monocytes using a recombinant human granulocytemacrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). Tumor cells were treated with LMB033 CPs containing a diarylethene fragment (photoswitch) in two ring forms - "closed" with low activity and toxicity and "open" with high activity. The obtained lysates of tumor cells were co-incubated with human monocyte-derived DCs. The analysis of the phenotypic characteristics of DCs was performed by a flow cytometry using monoclonal antibodies to CD83, CD86, CD11c, HLA-DR, and HLA-ABC. The expression level of mRNA of cytokine genes and indoleamine 2,3-dioxygenase (IDO) gene was determined using the quantitative real-time PCR.

Results: The highest cytotoxic effect on MDA-MB-231 cells was detected after 6-h incubation with the open form of LMB033 at concentrations of 16 and 32 μM. The studied CPs even at the lower of the tested concentrations caused externalization of phosphatidylserine in almost 100% of apoptotic cells of MB-MDA-231 cells following 6-h incubation. Loading monocyte-derived DCs with lysate of MDA-MB-231 cells treated with LMB033 peptide in open or closed forms caused a different effect on the antigen-presenting properties of cells depending on the form of the peptide. Compared to DCs loaded with untreated lysate, a significant increase in the number of mature activated CD83+ DCs was found after loading with lysates of cells treated with open (16 μM) or closed (32 μM) forms of LMB033. CPs-induced lysates of MDA-MB-231 cells did not cause significant changes in the expression of mRNA of Th1 polarizing cytokines TNF-α, IL-12, neither did these lysates activate the transcription of the genes of immunosuppressive cytokines and IL-10, TGF-β, and the IDO gene. This indicates the absence of the activation of the immunosuppressive properties of the generated DCs.

Conclusion: The presented data open the prospects for developing an effective antitumor immunotherapeutic vaccine based on DCs using CPs LMB033.

Background: The high heterogeneity and pathogenetic diversity of breast cancer (BCa) indicate the need for further study of tumor cell biology in order to identify molecular biological markers associated with the aggressiveness of tumors with a negative receptor status. Among many factors that may be involved in the initiation and progression of this form of cancer, the study of the immune components of tumor microenvironment, in particular PD-L1, is considered promising.

Aim: To investigate the relationship between PD-L1 expression in tumor tissue and clinical and pathological characteristics of BCa, taking into account the status of steroid hormone receptors.

Materials and methods: In tumor tissue of 116 patients with stage I-II BCa, the mRNA levels of CD274 gene were determined using the real-time quantitative polymerase chain reaction. The expression of PD-L1 was studied by the immunohistochemical method.

Results: The tissue of receptor-negative BCa was characterized by a significant decrease in the CD274 mRNA level against the background of the increased PD-L1 expression compared to neoplasms positive for the expression of steroid hormone receptors. An inverse correlation was found between PD-L1 at the protein level and the age of patients with receptor-negative BCa (r = -0.613, p = 0.00003). We showed that a characteristic feature of receptor-negative BCa in menopausal patients is the increased expression of PD-L1 both at the protein and mRNA levels (p = 0.005 and p = 0.046, respectively). The correlation of PD-L1 expression with metastatic lesions in regional lymph nodes (p = 0.050), tumor differentiation grade (p = 0.001), and the patient survival rate was revealed.

Conclusions: The obtained data indicated the expediency of using PD-L1 expression indicators for in-depth characterization of the tumor microenvironment of receptor-negative BCa, which will allow for the personalized correction of therapy regimens contributing to the improvement of the patients' quality of life.

Aim: To compare the expression of miRNA-185-5p and miRNA-424-5p in tumor cells and peripheral blood serum (PBS) of patients with endometrioid carcinoma of the endometrium (ECE) and to evaluate the significance of these biomarkers in cancer progression.

Materials and methods: The study was conducted on the samples of peripheral blood serum (PBS) and tumor tissue of 58 patients with stage I ECE using clinical and morphological methods and real-time polymerase chain reaction.

Results: A significant increase in the levels of circulating and tumor-associated miRNA-424-5p was established in ECE patients with a history of recurrences compared to patients without recurrences. To the contrary, the expression level of miRNA-185-5p increased in the PBS and decreased in the tumor tissue of ECE patients with recurrences compared to the patients without recurrence. In addition, we revealed that the expression levels of the studied miRNAs were associated with the differentiation grade and degree of tumor invasion. We established that miRNA-424-5p levels in PBS could serve as the most significant indicator for predicting the occurrence of recurrence in patients with ECE (AUC = 0.991; Sp 94.0%; Se 99.9%).

Conclusions: The expression features of miRNA-185-5p and miRNA-424-5p in the PBS and tumor tissue of patients with ECE are associated with the aggressiveness of cancer course and the risk of recurrence.

The study aimed to identify the clinically relevant gene variants in colon adenocarcinoma samples of Ukrainian patients using the NGS Comprehensive Cancer Panel (CCP) to implement them conveniently in clinical practice.

Methods: We have studied 20 samples of Ukrainian patients with colorectal adenocarcinomas of various differentiation grades. To identify the clinically relevant gene variants, the CCP data were filtered using the Franklin by Genoox database.

Results: A total of 79 clinically relevant gene variant alterations (SNVs, INDELs) were found in 28 of 409 genes. The largest number of mutations was found in 3 genes, APC, TP53, and KRAS (16, 14, and 8, accordingly). We revealed 4 variants in PTEN and SMAD4, 3 variants in CHEK2, ERBB2, and PIK3CA genes, and 2 variants in AKT1, ATM, DST, IDH1, and TCF12. Mutations for 7 genes, KRAS, TP53, CHEK2, PTEN, AKT1, APC, and SMAD4, were found in more than 1 tumor tissue sample. Tier 1-2 gene variants rate was about 50% of all genetic variants. The therapeutic significance was found in more than 55% of mutations. Additionally, 11 novel genetic mutations in 9 genes have been identified, including G6PD, APC, DST, SINE1, SMAD2, and FLCN.

Conclusions: These data suggest a high level of clinical relevance of the NGS CCP approach. Further confirmation on a larger number of samples and using a deeper analysis by other approaches is required.

Background: Imaging plays an important role in the identification of prostate cancer (PCa). However, a shortcoming of the current imaging techniques is their inability to detect PCa at an early stage of development when tumor volume is small. This led us to explore new and improved imaging methods. The phenomenon that infrared (IR) light penetrates biological tissues caused our efforts to utilize IR rays for PCa visualization. The aim of this study was to conduct model experiments to demonstrate how IR light could be used in the future to detect PCa in vivo.

Materials and methods: Experiments were carried out on prostates obtained after radical prostatectomy. The study was approved by the ethical commission of the Georgia-Israel Joint Clinic "Gidmedi". We developed a device that uses IR light to illuminate a prostate from the inside. In order to get IR images of the prostate, we developed a device with an IR-sensitive charge-coupled device (CCD) camera. The model experiments showed that the intensity of IR light passing through noncancerous and malignant prostate tissues is significantly different allowing their distinguishment. The visualization device can detect PCa lesions as small as several millimeters.

Conclusion: These results suggest that our device could be useful for the detection of small PCa lesions.

Background: The development of new approaches to modeling tumor radiosensitivity in patients with head and neck squamous cell carcinoma (HNSCC) is an important problem for overcoming tumor radioresistance. New agents for radiomodification are inhibitors of the enzyme cyclooxygenase-2 (COX-2). The study of markers of radioresistance in cancer patients undergoing radiotherapy (RT) in combination with COX-2 inhibitors and chemotherapy may contribute to the effectiveness of RT.

Aim: To determine the effect of conformal RT in combination with radiomodifiers (celecoxib, cisplatin, or their combination) on the content of vascular endothelial growth factor (VEGF), COX-2, and prostaglandin E-2 (PGE-2) in the serum of patients with HNSCC.

Materials and methods: 47 patients with HNSCC were divided into 4 groups: RT in combination with celecoxib and cisplatin, RT with cisplatin, RT with celecoxib, and RT. Patients received radiation treatment on a Clinac 600C linear accelerator. The levels of VEGF, COX-2, and PGE-2 in the serum were determined by enzyme immunoassay.

Results: Blocking COX-2 in patients with HNSCC leads to a decrease in VEGF levels. The largest decrease in VEGF levels was observed in a group treated by RT in combination with celecoxib and cisplatin, indicating a more effective antiangiogenic effect. The changes in the levels of VEGF, COX-2, and PGE-2, which are most pronounced under the combined effect of RT and both radiomodifiers, coincided with an objective response to radiation treatment.

Conclusions: The data obtained indicate the effect of radiomodification on the suppression of angiogenesis, which is most pronounced under the combined effect of RT and both radiomodifiers. The decrease in the levels of PGE- 2, COX-2, and VEGF coincides with the clinical efficacy of radiotherapy according to RECIST 1.1 criteria.