We studied interrelationships between the cytoarchitectonics of the mesenteric lymph node and the levels of microRNA-21, microRNA-221/222, and microRNA-429 in the lymph, blood serum, and breast tissues in female Wistar rats with chemically induced breast cancer. After polychemotherapy, significant correlations were found between miRNA-221 and the number of lymphoblasts in the germinal centers and between miRNA-222 and the number of lymphoblasts in the germinal centers and macrophages in the medullary cords of the mesenteric lymph nodes.

We performed a correlation analysis of the morphometry of mesenteric lymph nodes and cytokines of the thoracic duct lymph in chemically induced breast cancer (BC) and after surgical treatment of BC and neoadjuvant chemotherapy for BC according to the CMF scheme. After surgical treatment, correlations were found between the level of cytokines in the thoracic duct lymph and morphological changes in the mesenteric lymph nodes. The observed correlations can be a result of total resection of the tumor and increased immunomodulatory and antitumor action of cytokines. Comparison of the effect of chemotherapy and surgical treatment revealed correlations between cytokine concentrations in the lymph and structural changes in the lymph nodes, which can be related to weakening of local immunity and antitumor effect of cytokines.

We performed a morphometric analysis of mesenteric lymph nodes in rats with breast cancer induced by administration of N-methyl-N-nitrosourea. The volume of the paracortical zone and the number of mature plasma cells in the medullary sinuses were increased and the volume of lymphoid nodules with germinal centers and the number of macrophages were decreased in the group with tumor resection and chemotherapy in comparison with untreated rats with breast cancer. In rats receiving fragmented human double-stranded DNA in combination with adjuvant therapy, the volume of marginal and medullary sinuses and the number of small lymphocytes and macrophages in the paracortical zone increased in comparison with the group receiving chemotherapy without DNA preparation; the volume of lymphoid nodules with germinal centers returned to the level observed in the intact group; the volume of medullary substance and proliferative activity of cells in the germinal centers and medullary substance decreased, the number of mature plasma cells returned to normal in the medullary substance and decreased in the medullary sinuses.

Цель исследования - анализ корреляции морфометрии брыжеечных лимфатических узлов и концентрации цитокинов в лимфе грудного протока при химиотерапии рака молочной железы, хирургическом лечении и последующей химиотерапии. Методика. Рак молочной железы индуцировали введением N-метил-N-нитрозомочевины 5 раз с интервалом 7 сут подкожно в область 2-й молочной железы справа. Курс химиотерапии проходил по схеме CMF. Корреляцию между концентрациями 24 цитокинов лимфы и числом клеток структурных зон лимфатических узлов оценивали по коэффициенту ранговой корреляции Спирмена. Результаты. После химиотерапии РМЖ, по сравнению с РМЖ без лечения, морфологические преобразования в лимфатических узлах свидетельствуют о снижении активности местного иммунного ответа. Исследование корреляции концентрации цитокинов в лимфе со структурными изменениями в лимфатических узлах выявило зависимости направленные на повышение иммуномодулирующего и противоопухолевого действия цитокинов. После оперативного лечения РМЖ и последующей химиотерапии, по сравнению только с химиотерапией РМЖ, выявлены положительные связи иммунобластов с цитокином GRO/KC в герминативных центрах, цитокина IL-6 - с митотически делящимися клетками в герминативных центрах и мозговых тяжах, IL-5 - с иммунобластами в мозговых тяжах, хемокина MIP-1a - со зрелыми плазматическими клетками в мозговых синусах. Увеличено количество иммунобластов, средних и малых лимфоцитов в герминативных центрах, возросло количество малых лимфоцитов, незрелых и зрелых плазматических клеток в мозговых синусах. Увеличены площади мозговых тяжей и паракортикальной зоны. Выявлена корреляция: цитокина IL-1α с малыми лимфоцитами, IL-6 с иммунобластами, IL-7 и IL-18 - со средними лимфоцитами, GRO/KC - с иммунобластами, IL-17 - с макрофагами в Т-зависимой зоне; IL-7 и IL-18 - с иммунобластами, IL-12 - с макрофагами, MIP-1a и MCP-1 со зрелыми плазматическими клетками в мозговых синусах. Заключение. После оперативного лечения РМЖ c последующей химиотерапией, по сравнению только с химиотерапией РМЖ, выявлены взаимозависимости концентрации цитокинов в лимфе грудного протока с морфологическими изменениями в брыжеечных лимфатических узлах, которые могут указывать на повышение активности местного звена иммунного ответа. The aim of this study was to analyze correlations of the morphometry of mesenteric lymph nodes with cytokine concentrations in thoracic duct lymph in chemotherapy and surgical treatment with subsequent chemotherapy of breast cancer. Methods. Breast cancer was induced by subcutaneous injection of N-methyl-N-nitrosourea 5 times with 7-day intervals, into the region of the 2nd breast on the right. The course of chemotherapy was performed according to the CMF scheme. Correlations between concentrations of 24 cytokines of the lymph and cells of lymph node structural regions were estimated by the Spearman rank correlation coefficient. Results. After chemotherapy for breast cancer compared to untreated breast cancer, morphological transformations in lymph nodes indicated decreased activity of the local immune response. Analysis of correlations between lymph concentrations of cytokines and structural changes in lymph nodes identified relationships aimed at increasing the immunomodulatory and antitumor effects of cytokines. After surgical treatment of breast cancer and subsequent chemotherapy compared to chemotherapy alone, positive correlations were found for immunoblasts with cytokine GRO/KC in germinative centers, for cytokine IL-6 with mitotically dividing cells in germinative centers and medullary cords, for IL-5 with immunoblasts in medullary cords, and for chemokine MIP-1a with mature plasma cells in medullary sinuses. Numbers of immunoblasts and medium and small lymphocytes were increased in germinative centers whereas numbers of small lymphocytes and immature and mature plasma cells were increased in medullary sinuses. Areas of medullary cords and the paracortical zone were increased. Correlations were found for cytokine IL-1α with small lymphocytes, for IL-6 with immunoblasts, for IL-7 and IL-18 with medium lymphocytes, for GRO/KC with immunoblasts, for IL-17 with macrophages in the T-dependent zone, for IL-7 and IL-18 with immunoblasts, for IL-12 with macrophages, and for MIP-1a and MCP-1 with mature plasma cells in medullary sinuses. Conclusion. After surgical treatment of breast cancer and subsequent chemotherapy compared to chemotherapy alone, cytokine concentrations in lymph of the thoracic duct were observed to correlate with morphological changes in mesenteric lymph nodes, which may indicate increased activity of the local immune response.

We performed a correlation analysis of the morphometric parameters of mesenteric lymph nodes and cytokine content in the lymph of thoracic duct in rats with chemically induced breast cancer. The study showed that activity of the local immune response in the lymph nodes in breast cancer is aimed at antitumor protection. In breast cancer, the area of the paracortical zone remained at the level of the intact group, while the area of lymphoid nodules with germinative centers and the area of medullary substance increased; the number of macrophages in the thymus-dependent zone and zone responsible for humoral immunity also increased. The following positive correlations were revealed: in germinative centers and medullary substance, number of mitotic cells correlated with cytokine IL-5 content and the number of medium lymphocytes correlated with the content of chemokine MIP-1α; in the germinative centers, the number of immunoblasts correlated with the level of cytokine GRO/KC, in the paracortical zone, the number of macrophages correlated with the level of chemokine MCP-1, the number of reticular cells correlated with IL-6 and M-CSF content; in medullary substance, the number of small lymphocytes and mature cells plasma cells (their content was reduced) correlated with the level of chemokine GRO/KC, which can be caused by their migration from the lymph node.

We studied the levels of serum and lymph cytokines involved in the pathogenesis of BC. BC was induced by injection of N-methyl-N-nitrosourea to Wistar rats. The animals underwent surgery, or received polychemotherapy (cyclophosphamide, methotrexate, and 5-fluorouracil), or surgical treatment was combined with polychemotherapy; in a special group, Panagen (fragmented DNA) was added to polychemotherapy. Cytokine concentration in the lymph was measured using Bio-Plex Pro Rat Cytokoness 24-Plex Assay test system (Bio-Rad). In rats with BC, the content of most studied cytokines (IL-1β, IL-2, IL-4, IL-6, IL-7, IL-12, IL-13, MIP-1α, MIP-3α, RANTES, TNFα, and MCP-1) in the lymph and blood was significantly higher, while the content of IL-10 and GRO/KC was lower than in intact animals. Surgical resection of the tumor led to a significant decrease in the content of both pro- and anti-inflammatory cytokines in the lymph. Polychemotherapy led to a significant decrease in the content of IL-1β, IL-4, IL-6, IL-7, MIP-1α, MIP-3α, and RANTES in the serum and lymph. Comparison of the cytokine content in the serum and lymph of operated animals after polychemotherapy with and without Panagen showed that the content of most cytokines (IL-5, IL-6, IL-7, IL-10, IL-13, IL-17A , IL-18, GRO/KC, IFNγ, and MIP-3α) was higher after Panagen administration.

The morphometric analysis of mesenteric lymph nodes was carried out in female Wistar rats with chemically induced breast cancer. In control rats with untreated breast cancer, the volume of the system of sinuses increased in parallel with the appearance of morphological signs of suppression of cell-mediated immunity, inhibition of humoral immunity, and macrophage reaction. Against the background of chemotherapy, we observed a decrease in the volume of paracortex and lymphoid nodules, suppression of proliferative activity of lymphoid cells in paracortical and B-cell zone, and a decrease in macrophage content. After resection of breast cancer followed by chemotherapy course, lymph transport activation, widening of the paracortex, enhanced proliferative activity of cells in the paracortex and B-cell zone, and reduced volumes of lymphoid nodules with and without germinal centers and medullary substance were revealed in comparison with rats subjected neoadjuvant chemotherapy.

The effects of various treatment modes on the morphology of anterior mediastinal lymph nodes were examined in female Wistar rats with chemically provoked breast cancer. Adjuvant chemotherapy impaired filtration barrier potential of the anterior mediastinal lymph nodes, which manifested in increased volume of sinuses, reduced volumes of lymphoid nodules with germinal centers and thymus-dependent regions, down-regulated proliferative activity of lymphoid cells in B-cell zone and paracortex, and diminished macrophage score in all zones. Intraperitoneal injection of double-stranded DNA preparation (5 mg/kg) activated the humoral and cellular immune responses manifested by morphological alterations in anterior mediastinal lymph nodes observed in parallel with a decrease of medullary sinuses volume: enhancement of lymphocyte volume and lymphocyte score in paracortex, mantle zone expansion, and an increase of volume of the light centers in lymphoid nodules paralleled with diminished proliferative activity in them.

The histological and immunohistochemical type of chemically induced (injection of N-methyl-N-nitrosourea into the mammary gland) breast tumor was studied in Wistar females. The tumor induced by N-methyl-N-nitrosourea was moderately differentiated adenocarcinoma with infiltrative growth lacking estrogen-α and human epidermal growth factor receptors, and expressing progesterone receptors; tumor cells were characterized by high proliferative activity. This variant of chemically induced breast tumor corresponds to human breast cancer luminal type B.

Histological study of structural transformations in the thymus of Wistar females in induced carcinogenesis (N-methyl-N-nitrosourea injection in the right 2-nd mamma) and polychemotherapy (6 months after tumor growth initiation; cyclophosphamide, methotrexate, and 5-fluorouracyl) was carried out. The area of the cortical matter in the thymus decreased 6 months after carcinogenesis induction, the percentage of connective tissue elements and glandular tissue and the counts of immunoblasts and cells with pyknotic nuclei increased, this indicating the development of accidental involution of the thymus. Animals of the experimental tumor+chemotherapy group exhibited morphological signs of lymphocyte migration from the thymus and suppressed activities of the lymphoid and epithelial components (lesser area of connective tissue elements and glandular tissue, lesser density of parenchymatous cell elements, lesser counts of immunoblasts and small lymphocytes, and larger area of the medulla) in comparison with animals without chemotherapy.

We studied hormone levels in the thoracic duct lymph and expression of miRNA involved in the pathogenesis of breast cancer induced in rats by intramammary injection of N-methyl-Nnitrosourea. The correlations between miRNA expression and hormone levels depended on the type of treatment.

The anterior mediastinal lymph nodes were analyzed morphometrically in rats with chemically provoked breast cancer. Rats with untreated breast cancer and animals receiving chemotherapy demonstrated decreased volumes of paracortical region and lymphoid nodules with the germinal centers accompanied by extended medullary thymic substance. Resection of largest focus of breast tumor improved the filtration barrier potential of anterior mediastinal lymph nodes, up-regulated the proliferative activity of lymphoid cells in T-cell zones, and down-regulated proliferation of plasmatic cells.

Morphological changes in the thymus of female Wistar rats with experimental mammary gland carcinomas were studied. After adjuvant therapy, the area of the cortical matter and density of parenchymal cells in the thymus decreased, while areas of the medulla, connective tissue, and content of immunoblasts and macrophages increased. In the thymuses of rats receiving exogenous DNA, morphological signs of activation of the lymphoid and epithelial components were found: areas of the cortex and medulla, glandular and connective tissue corresponded to the values in intact animals, the counts of lymphocytes in the central part of the cortical matter and of macrophages in all zones of the thymus increased, and lymphocyte migration from the thymus increased (in comparison with the chemotherapy group).

The work purpose was to reveal an existence of an associativity of the microRNA levels in blood serum to quantitative and functional indices of cells haemo - and lymphopoiesis at the experimental breast cancer induced by N -methyl - N- nitrosourea in the remote period after surgery and carrying out neoadjuvant polychemotherapy. At animals there were investigated levels of microRNA-21, microRNA-221, microRNA-222 and microRNA-429 in serum, also investigated quantitative and functional parameters of cells from bone marrow, from lymph of a chest channel and from spleen. Statistically significant distinctions on the microRNA level in blood serum and an existence of interrelations of microRNA levels with quantitative and functional indices of haemo- and lymphopoiesis cells were revealed.

The involvement of hormones, microRNAs and cytokines in breast cancer pathogenesis has been well established. Lymph picks up secretory products of breast cancer cells. The purpose of the study was to evaluate the levels of hormones, microRNAs and cytokines in lymph. Wistar rats were injected with N-methyl-N-nitrosourea to induce breast cancer. The rats were subjected to either surgery alone or chemotherapy alone (cyclophosphamide, methotrexate and 5-fluorouracil). In some animals, surgery was followed by chemotherapy. The levels of follicle-stimulating hormone (FSH), prolactin, luteinizing hormone (LH), estradiol (E2) and thyroglobulin (TG), microRNA-21, microRNA-221, microRNA-222, microRNA-429 and 24 cytokines were determined. Chemotherapy was shown to result in the reduction in the levels of prolactin, thyroglobulin, FSH and estradiol. In rats with breast cancer, the expression levels of microRNA-21, microRNA-221 and microRNA-222 were increased, and the expression levels of microRNA-429 were decreased. In breast cancer rats, the levels of most cytokines were found to be increased. Correlations between the levels of cytokines, hormones, and microRNAs in lymph were identified. Differences in the expression levels of cytokines, hormones, and microRNAs in lymph with respect to treatment option were detected.

In work the microRNA levels in tissues of a mammary gland at experimental model of a breast cancer at rats of the Wistar are investigated. The levels of oncogenic microRNA (21, 221, and 222) in tumor tissues are increased, and the level of tumor-suppressive microRNA (429) are decreased. Under the influence of neoadjuvant polychemotherapy suppression of levels of an expression the oncogenic microRNA and increase in levels of expression microRNA tumor-suppressive is noted.

In work was done the analysis of microRNA expression breast tissue and their levels in the blood and lymph in normal and breast cancer. It is shown that expression of microRNA in tumor tissue and levels in blood and in lymph was elevated. The relationships between the expression levels of the breast tissue microRNAs with their content in the blood and lymph, as well as the type of the treatment was established.

In the pathogenesis of diseases, including cancer, changes in the levels of expression of microRNAs play an essential role. The aim of the research was to study the levels of microRNA in the lymph in experimental breast cancer and correlation with the number and functional activity of cells of the hemo – and lymphopoiesis. Breast cancer induced by the introduction of n-methyl-N-nitrosourea in Wistar rats. In the lymph investigated the content of microRNA-21, 221, 222, 429. It is shown that between the levels of expression of tissue breast tumor microRNA, the number and functional activity of cells of the hemo – and lymphopoiesis were present a some correlations. The levels of microRNAs in breast cancer depend on the type of treatment and are associated with parameters of cells hemo – and lymphopoiesis.