A review of the literature data on expression of estrogen receptor alpha and beta (ERa and ERβ) in tumors different from breast cancer. The results regarding the ERa and ERβ expression frequency in non-small cell and small cell lung cancer, colorectal cancer, esophageal, ovarian, prostate and brain tumors are presented. High frequency of estrogen receptor expression (in up to 50 and more per cent of cases) in various types of tumors, differences between ERa and ERβ in expression frequency, prognostic significance and prediction of the neoplastic process aggressiveness as well as in biological implications of interaction with antiestrogens (antagonistic and/or agonistic effect) are shown. The data on comparative evaluation of ERa and ERβ expression in lung, ovarian, prostate tumor cells and corresponding nonneoplastic tissues are reported. Authors consider necessary to include the ERa and ERβ detection into the routine clinical practice not only in breast cancer but in other tumors as well. Prospects of the clinical application of antiestrogens, in particular tamoxifen, in adjuvant therapy of different tumors with positive ER status are discussed.

Objectives. Quantitative assessment of the expression and co-expression levels of estrogen receptors of different types (ERα and ERβ) in malignant tissues of large cohort of patients with non-small cell lung cancer (NSCLC). Materials and methods . Measurement of ERα and ERβ expression levels was carried out on 167 malignant tissue samples by immunofluorescence flow cytometric method. Level of the ERα and ERβ expression was calculated as a ratio of specifically fluorescent cells (%) compared to the control (incubated with the secondary antibody only). Results. The expression of ERα and ERβ was detected in all the NSCLC samples investigated. High variability in the level of the marker expression was revealed and it was shown 1,5–2,0-fold higher ERβ expression in the most tumors than that for ERα. The Spearman rank correlation coefficient of 0,364 (p <0,001) and the Pearson correlation coefficient of 0,401 (p <0,001) indicate statistically significant weak positive correlation between the levels of expression of ERβ and ERα. Conclusion . The level of expression of ERβ as a main subtype of ER in NSCLC tissue could prove useful for selecting patients for antiestrogen therapy

Non-degradable steel and titanium implants used to replace defects of the locomotor system or fabricate vascular stents provide maximum stability but have too many drawbacks. Currently, biodegradable magnesium alloys are considered as promising materials for creation of fixation devices in orthopedics and cardiovascular surgery. First attempts of using magnesium-based implants for bone fixation were made as early as at the beginning of the 20th century, however, due to a high corrosion rate and gas formation they turned out to be unsuccessful. Magnesium-based alloys developed recently demonstrate improved anti-corrosion and mechanical properties and are promising for manufacturing of biodegradable, biocompatible metal implants. The microstructure of magnesium implants, their mechanical properties, electrochemical behavior, and kinetics of degradation are affected by alloying elements, methods of surface coating, and thermomechanical treatment of implants. All these factors determine the rate of alloy degradation in physiological environment and the level of gas formation. Although preclinical studies and even singular pilot clinical trials of the medical devices based on magnesium alloys have been carried out recently, there remain many unsolved issues preventing the introduction of biodegradable magnesium alloys in clinical practice. This review discusses the most promising directions in the development of biomedical materials based on magnesium alloys, existing limitations, and challenges of their use. The possibility of employing biodegradable magnesium alloys in oncology is also shown.

Estrogens play significant role in development and progression of non-small cell lung cancer (NSCLC), but prognostic value of estrogen receptors (ERα and ERβ) in cancer tissue are controversial. Moreover, there is no accurate information about frequency of ERs expression in tumor cells as the key for antiestrogens which is a promising NSCLC therapy. Therefore, we aimed to 1) evaluate expression of ERβ in NSCLC cancer tissue and 2) estimate prognostic value of ERβ on survival of patients. Expression of ERβ in 125 surgical NSCLC samples was evaluated quantitatively by immunofluorescent assay and flow cytometry. The primary anti-ERβ (14C8, ab288) and secondary (DyLight650, ab98510) antibodies were used. The level of ERβ expression was calculated as the ratio (%) of specifically fluorescent cells to the control (incubation with the secondary antibody only). The prognostic value of ERβ expression in tumor tissue was studied in 49 patients without neoadjuvant therapy who died in 6.5 years after radical surgery. There was division into two level expression groups: high – ERβ were revealed in ≤40% and low – >40% cells. Statistical analysis was performed by Kaplan-Meier curves analysis (log-rank test) and Cox-regression in software packages: GraphPad Prism 7.0 and SPSS 22.0. 1. Expression of ERβ was revealed in all the investigated NSCLC samples. Significant heterogeneity of the marker expression level was shown: the mean and median of the marker index were 42.3%±15.6 and 44.0% respectively. 2. It was shown in comparison of Kaplan-Meier curves in patients with low vs high level of ERβ expression (see Fig.1) that the survival median of the patients who died in 6.5 years after radical surgery was more than 1.5 times greater in the group with the low vs high level of ERβ expression: 36.0 and 22.0 months respectively (p=0.04). The risk of death in follow-up period after radical surgery was 2.0 times greater in the group classified as high level of tumor ERβ expression: hazard ratio has been statistically significant – 1.8 (95% CI: 1.1–6.4, p=0.05). 1. ERβ tumor expression has prognostic value in NSCLC patients, namely, the high level of ERβ (in more than 40% tumor cells) is a negative prognostic marker. 2. The fact of ERβ expression in all the NSCLC specimens investigated has shown that hormonal antiestrogen therapy could be a promising option for all the NSCLC patients.

The differences in expression of ERCC1 were estimated between tumor specimens embedded into paraffin blocks and surgical biopsy specimens of non-small cell lung cancer as well as breast and ovarian cancers. Concordance or differences not higher than 20% were observed in 73% of the cases. The number of the cases with more significant differences in ERCC1 expression was less than 17%. The results show that ERCC1 detection in surgical biopsy specimens by flow cytometry is the more preferable method due to reduced preanalytical phase of the analysis.

Background: Estrogen receptors beta (ERβ) are highly expressed in different normal and neoplastic tissues that, until recently, has been considered to be ER-negative based on ERα expression evaluation. Among the genes, regulated via estrogen signaling there is one, coding microtubule protein beta-III tubulin (TUBB3). TUBB3 expression is found in many solid tumors and is linked to poor prognosis and resistance to taxanes. Since it is little known about mechanisms behind TUBB3 expression in non-small cell lung cancer (NSCLC), we decided to find out if there is a correlation between ER and TUBB3 expression in this type of cancer. Methods: 104 surgical samples of NSCLC were converted to single-cell suspension, stained with primary anti-ERα (abSP-1), anti-ERβ (ab14C8), anti-TUBB3 (ab7751) antibodies and secondary fluorescent antibodies. Immunofluorescent estimation was performed using flow cytometry. Expression level was determined as the ratio (%) of specifically fluorescent cells to the number of cells stained with secondary antibodies. Spearman rank correlation was used to test the association between variables. Results: Both ER were revealed in all NSCLC specimens. Mean expression level of ERβ was significantly higher compared with ERα (46,6 ± 17,0% vs 23,2 ± 14,2%, respectively). Mean TUBB3 expression level was 43,1±15,7%. In all the tumors investigated only weak correlation observed between and ER status and TUBB3 expression level (rs = 0,3 and rs = 0,4 for ERα and ERβ, respectively). In the group of squamous cell cancer specimens (n = 68) the association was strong (rs = 0,5 and rs = 0,5 for ERα and ERβ, respectively). In the group of adenocarcinoma specimens (n = 36) the correlation between ERα and TUBB3 was very weak (rs = 0,3) and there was no correlation between ERβ and TUBB3. Conclusions: 1. Strong correlation between TUBB3 and ER expression was found only in squamous cell cancer tissue. 2. The dominant type of estrogen receptors is ERβ. 3. In clinical terms high ERβ expression means that in case of resistance to standart platinum/taxane duplets, patients with high tumor ERβ expression may benefit from antiestrogen therapy. Supported by RFBR grants (№№15-04-06991-а, 16-34-01049-mol-a) and grant of the President of RF МК-7709.2016.7. Legal entity responsible for the study: N.N. Blokhin Russian Cancer Research Center. Funding: Russian Foundation for Basic Research Grants, grant of the President of Russian Federation. Disclosure: All authors have declared no conflicts of interest.

Background. Epithelial-mesenchymal transition (EMT) is a factor related to metastatic potential of tumor cells. The most distinguishing feature of this transformation is expression of protein vimentin, which is not common for epithelial cells. Data of EMT level and clinical significance of the marker is ambiguous in prognosis of tumor different localization including ovarian cancer. Objective is characterization of EMT in ovarian cancer tissue based on quantitative analysis of co-expression level of epithelial cytokeratins and mesenchymal cells marker vimentin. Materials and methods. A quantitative assessment of co-expression level of cytokeratins and vimentin was performed by double immunofluorescence staining method (58 surgery specimens of ovarian cancer stage III), associated with flow cytometry. Results. Double immunofluorescence staining method, developed and used in the current study, was used for quantitative assessment of EMT level based on value of cytokeratin and vimentin co-expression in epithelial cells. Epithelial cells co-expressed these markers, were detected in 100 % tumor investigated with individual value differences from 10 to 86 %. Average co-expression level of cytokeratins and vimentin in subgroups with high and low level value related to median 42 % significantly varied and were 28,5 ± 7,5 and 56,3 ± 11,8 % (p = 0,02) respectively. Conclusions Serous ovarian cancer is molecular heterogeneous group with principal differences in level of EMT tumor phenotype between various patients. In half of the cases the disease is characterized by high metastatic potential of tumor cells. Co-expression of cyto-keratins and vimentin in all the tumors investigated is evidenced clinical significance of this molecular characteristic in ovarian cancer pathogenesis.

Background. A search for pathogenetically grounded approaches to the treatment of non-small cell lung cancer is undoubtedly one of the top priorities, because of the results of its treatment cannot be regarded as satisfactory. Discovery of a new type of receptors, estrogen receptors ß, that expressed in non-small cell lung cancer, created the preconditions for the development of a new treatment strategy for non-small cell lung cancer, namely antiestrogen therapy. Objective was to answer to the clinically significant question of how many patients with non-small cell lung cancer and lung metastases are potential candidates for antiestrogen treatment. Materials and methods. A comparative quantitative estimation of the level and frequency of the estrogen receptors ß expression in non-small cell lung cancer tissues and also in lung metastases (74 in total) was carried out by flow cytometry. Results. The estrogen receptors ß expression was detected in the majority of the investigated tumors and lung metastases, in 92 and 86 % of patients, respectively. The average level of estrogen receptors ß expression in non-small cell lung cancer tissue was higher then in metastases (42 % and 34.6 % respectively). The differences was statistically significant (p = 0.03). Primary tumors with a high and low+moderate estrogen receptors ß expression levels were detected in 35 % and 65 % of cases respectively and metastases with such expression levels - in 14 % and 86 % of cases respectively. Conclusion. 1. The estrogen receptors ß expression in the tumor predicts a more favorable course of the disease. One of the reasons for this may be a greater metastatic potential of tumor cells with low estrogen receptors ß expression. 2. The group of potential candidates for carrying out the adjuvant antiestrogen therapy comprises about 70 % of patients with primary lung tumors and with metastatic lesions of lung.

Background. Beta-III tubulin (TUBB3) is a tumor-specific isoform of the microtubule protein beta-tubulin. TUBB3 is considered to be a marker of adverse prognosis and tumor resistance to therapy with taxanes and Vinka alkaloids. Association between TUBB3 expression and histological type of the tumor has not been studied properly yet. Objective. The expression level of TUBB3 in non-small cell lung cancer biopsy specimens has been measured on 2 groups of patients with adenocarcinoma and squamous cell carcinoma. Materials and methods. The samples with adenocarcinoma (n = 43) and squamous cell carcinoma (n = 39) were converted to suspension, filtered, fixed with 4 % formaldehyde, stained with monoclonal antibodies for TUBB3 and DyLight 650-conjugated secondary antibodies to mouse IgG and analyzed by flow-cytometry. The method was developed in N.N. Blokhin Russian Cancer Research Center. Results. The average level of TUBB3 expression in the adenocarcinoma group is higher than in the squamous cell carcinoma group (33.1 ± 12.4 % of cells expressing the marker in adenocarcinoma vs. 26.0 ± 13.6 % in squamous cell carcinoma; differences were statistically significant). The average level of TUBB3 expression in adenocarcinoma is 29.7 ± 8.1 % in female and is 34.9 ± 13.9 % in male (differences statistically insignificant). Since the group of patients with adenocarcinoma was presented by both men and women, while the group of patients with squamous cell carcinoma was only men, from the analysis was excluded all female patients. Differences between the groups remained statistically significant. Conclusion. TUBB3 expression in tumor tissue does not depend on the gender, at least among patients with adenocarcinoma of the lung, and at the same time, the level of TUBB3 in adenocarcinoma tissue is higher in comparison with squamous cell carcinoma tissue.

Using the model of breast cancer Ehrlich ascites tumor in mice, we showed that a sigle intraperitoneal injection of cardiac glycoside digoxin 1 h before the intraperitoneal injection of cisplatin increased the anticancer effect of the cytostatic drug more than twice when recalculated for the dose. It is assumed that the modifying effect of digoxin is determined by the direct inhibition of glycolysis in tumor cells. Taking into account the design of the study, we consider promising the clinical evaluation of the effectiveness of digoxin as a modifier of cisplatin efficiency in intracavitary therapy of ascites cancers with pleural and abdominal dissenmination.

Tamoxifen is the first target agent with a high-end position in breast cancer therapy till now. In recent years experimental researches revealed new biological effects of tamoxifen on tumor cells. The present study continues the theme of the review published in 2012, where a plenty of tamoxifen effects besides interaction with estrogen receptors was discussed. Thus, there is described a wide range of the drug targets which are the key points of signal cascades activating the cell proliferation and determining the course of the growth of the cancer and its sensitivity to chemotherapy. Also clinical trials of tamoxifen based on existing of targets besides the estrogen receptors are reviewed. Furthermore, the data on the antiviral, antibacterial, antifungal and antiparasitic activities of tamoxifen are indicated.

Background. Class III beta-tubulin (TUBB3) is one of the eight beta tubulin isotypes identified in human. It is constitutively expressed in brain and testis but not in lung tissue. TUBB3 is also known to appear in solid tumors, in particular in non-small cell lung cancer (NSCLC), and it is often associated with poor prognosis and resistance to taxanes and Vinka alkaloids. Objective: We have suggested that TUBB3 expression may cover not only the primary tumor but also a wide adjacent area of morphologically normal lung tissue. To check the hypothesis we have measured TUBB3 expression both in the non-small cell lung carcinoma (NSCLC) and remote lung tissue derived from the same lung. 60surgical biopsy specimens of NSCLC and morphologically normal lung tissue of 30patients were investigated. Materials and methods. Biopsy specimens were converted to suspension, fixed in 4 % formaldehyde and analyzed by flow cytometry using monoclonal antibodies to TUBB3. The method was developed in the laboratory of medicinal chemistry research Institute of experimental diagnostics and therapy of tumors (N.N. Blokhin Russian Cancer Research Center) and patented. Results. Fraction of TUBB3-positive cells in the group of adjacent lung tissue specimens was lower compared to NSCLC group but still positive in the majority of adjacent lung tissue specimens (25 of 30) and achieved up to 39 % of cells. Conclusion. We suggest that TUBB3 expression in adjacent morphologically normal lung tissue indicates presence of transformed and potentially malignant cells far from the primary site.

T.A. Bogush1, I.A. Mamichev1, E.A. Dudko1, A.N. Grishanina1, R.J. Ramanauskaite1, N.O. Vichljantzeva1, S. Tjulandin2, B.E. Polotsky3, M.I. Davydov3 Laboratory of Medical Chemistry, N. N. Blokhin Russian Cancer Research Center, Moscow, Russian Federation Department of Clinical Pharmacology and Chemotherapy, N. N. Blokhin Russian Cancer Research Center, Moscow, Russian Federation Surgical Department, N. N. Blokhin Russian Cancer Research Center, Moscow, Russian Federation

Aim: Beta–III tubulin or TUBB3 is normally expressed in neuronal cells and testicular Sertoli cells only. However, TUBB3 has been revealed in solid tumors and high levels of the marker is associated with poor disease prognosis. We have suggested that TUBB3 expression in morphologically normal tissue adjacent to the tumor might be an additional prognostic marker which indicates a malignization around the tumor. To check the hypothesis we have compared quantitatively TUBB3 expression in non-small-cell lung carcinoma (NSCLC) with its expression in adjacent lung tissue.

Aim: The results of cancer chemotherapy are not satisfactory enough; that is why a search of pathogenic based personalized approaches is a top priority task of oncology. ER beta creates a new foundation for this. We have revealed high levels and frequency of ER beta in non-small cell lung cancer (NSCLC) and proposed this marker as a target for adjuvant antiestrogen therapy of the disease. In the study we have answered the questions 1) if this approach is appropriate for metastatic lung cancer and 2) could the comparison of ER beta expression in primary and metastatic lung cancer support a good prognostic value of the marker?Methods: 83 surgical specimens of NSCLC and lung metastasis were analyzed by flowcytometry after incubation with primary (clone 14C8, ab288, Abcam) and secondary (F2772, Sigma) antibodies. Mean cell fluorescence and number of stained cells were analyzed with WinMDI software and Kolmogorov-Smirnov approach. Three level of ER beta were analyzed: high – ER was revealed in more than i50% of the cells; moderate – in 30-49%; low – in less than 30%.Results: 1. ER beta was revealed in most of the primary (92%) and metastatic (86%) tumors with lower mean level of expression in metastases (42% and 34%, р = 0.03) and with no differences in mean ER beta intensity (р = 0.06). 2. High ER beta level was revealed in 35% of NSCLC, but in metastases – in 14 % (p = 0.03). 3. The cases with moderate plus low ER beta level were revealed in 65% of NSCLC, but in metastases the index was higher – 86% (p = 0.03). 4. Number of cases with ER beta level more than 30% (moderate plus high level) was similar in NSCLC and metastases (73 vs. 65%, p = 0.08).Conclusions: 1. Lower indexes of ER beta expression in lung metastases cells with high metastatic potential compared with primary lung tumors confirm the good prognostic value of ER beta in NSCLC. 2. More than half of lung metastasis patients, as well as about 70% of patients with NSCLC with high and moderate level of ER beta expression, could benefit from adjuvant antiestrogen therapy. Supported by RFBR grants (13-04-01004-a, 15-04-06991-a, 14-04-31734–mol-a), FIMT-2014-205 and scholarship of the President of Russian Federation (376.2012.4).Disclosure: All authors have declared no conflicts of interest. Aim: The results of cancer chemotherapy are not satisfactory enough; that is why a search of pathogenic based personalized approaches is a top priority task of oncology. ER beta creates a new foundation for this. We have revealed high levels and frequency of ER beta in non-small cell lung cancer (NSCLC) and proposed this marker as a target for adjuvant antiestrogen therapy of the disease. In the study we have answered the questions 1) if this approach is appropriate for metastatic lung cancer and 2) could the comparison of ER beta expression in primary and metastatic lung cancer support a good prognostic value of the marker? Methods: 83 surgical specimens of NSCLC and lung metastasis were analyzed by flowcytometry after incubation with primary (clone 14C8, ab288, Abcam) and secondary (F2772, Sigma) antibodies. Mean cell fluorescence and number of stained cells were analyzed with WinMDI software and Kolmogorov-Smirnov approach. Three level of ER beta were analyzed: high – ER was revealed in more than i50% of the cells; moderate – in 30-49%; low – in less than 30%. Results: 1. ER beta was revealed in most of the primary (92%) and metastatic (86%) tumors with lower mean level of expression in metastases (42% and 34%, р = 0.03) and with no differences in mean ER beta intensity (р = 0.06). 2. High ER beta level was revealed in 35% of NSCLC, but in metastases – in 14 % (p = 0.03). 3. The cases with moderate plus low ER beta level were revealed in 65% of NSCLC, but in metastases the index was higher – 86% (p = 0.03). 4. Number of cases with ER beta level more than 30% (moderate plus high level) was similar in NSCLC and metastases (73 vs. 65%, p = 0.08). Conclusions: 1. Lower indexes of ER beta expression in lung metastases cells with high metastatic potential compared with primary lung tumors confirm the good prognostic value of ER beta in NSCLC. 2. More than half of lung metastasis patients, as well as about 70% of patients with NSCLC with high and moderate level of ER beta expression, could benefit from adjuvant antiestrogen therapy. Supported by RFBR grants (13-04-01004-a, 15-04-06991-a, 14-04-31734–mol-a), FIMT-2014-205 and scholarship of the President of Russian Federation (376.2012.4). Disclosure: All authors have declared no conflicts of interest.

A replacement of major defects of the trachea remains a challenge despite numerous attempts to use for this purpose various options of autologous and allogeneic implants as well as synthetic matrixes. The most prospective direction is to create tracheal matrixes based on biocompatible porous or fibrous materials that mimic native tissues and provide the proliferation of multipotent cells. This review summarizes current data regarding the development of experimental research and clinical testing of biocompatible tracheal matrixes, which were created using innovative technologies.