Целью работы явилась оценка связи хромосомных аберраций генов BRCA1, NF-κB1, PARP1 в опухолевой ткани молочной железы с эффектом химиотерапии и прогнозом заболевания. В исследование включены 85 больных люминальным В раком молочной железы IIA-IIIB стадии, с морфологически верифицированным диагнозом, в возрасте 25-68 лет (47,8±1,1). Было установлено, что наибольшая частота делеций наблюдается в гене BRCA1 (36%, 30 случаев из 85). Частота амплификаций PARP1 в исследуемой группе больных РМЖ достигает 62% (53 случая из 85). Наименьшее число хромосомных аберраций наблюдается в гене NF-κB1, всего у 22 больных наблюдаются делеции и амплификации данного гена (26%). Показано, что делеция гена BRCA1 сопряжена с хорошим ответом на неоадъювантную химиотерапию вне зависимости от наличия хромосомных аберраций других генов. При этом амплификация PARP1 связана с плохим прогнозом заболевания. The aim of the work was to assess the relationship of chromosomal aberrations of the BRCA1, NF-κB1, PARP1 genes in breast tumor tissue with the effect of chemotherapy and prognosis of the disease. The study included 85 patients with luminal stage IIA-IIIB breast cancer, with a morphologically verified diagnosis, aged 25-68 years (47.8±1.1). As a result of the study, the CNA frequency of the studied genes was estimated. It was found that the highest deletion rate is observed in the BRCA1 gene (36%, 30 cases out of 85). The frequency of amplification of PARP1 in the studied group of breast cancer patients reaches 62% (53 cases out of 85). The smallest number of chromosomal aberrations is observed in the NF-κB1 gene; in only 22 patients, deletions and amplifications of this gene are observed (26%). it was found that the deletion of the BRCA1 gene is associated with a good response to neoadjuvant chemotherapy, regardless of the presence of chromosomal aberrations of other genes. Moreover, amplification of PARP1 is associated with a poor prognosis of the disease.

The aim of the work was to assess the relationship of chromosomal aberrations of the BRCA1, NF-κB1, PARP1 genes in breast tumor tissue with the effect of chemotherapy and prognosis of the disease. The study included 85 patients with luminal stage IIA-IIIB breast cancer, with a morphologically verified diagnosis, aged 25-68 years (47.8±1.1). As a result of the study, the CNA frequency of the studied genes was estimated. It was found that the highest deletion rate is observed in the BRCA1 gene (36%, 30 cases out of 85). The frequency of amplification of PARP1 in the studied group of breast cancer patients reaches 62% (53 cases out of 85). The smallest number of chromosomal aberrations is observed in the NF-κB1 gene; in only 22 patients, deletions and amplifications of this gene are observed (26%). it was found that the deletion of the BRCA1 gene is associated with a good response to neoadjuvant chemotherapy, regardless of the presence of chromosomal aberrations of other genes. Moreover, amplification of PARP1 is associated with a poor prognosis of the disease.

Страница 82 МЕТАБОЛИЧЕСКАЯ ВИЗУАЛИЗАЦИЯ РАКА МОЛОЧНОЙ ЖЕЛЕЗЫ МЕТОДОМ ОДНОФОТОННОЙ ЭМИССИОННОЙ КОМПЬЮТЕРНОЙ ТОМОГРАФИИ С 99M TC

Molecular imaging is a multimodal discipline for visualizing biological processes at the subcellular level in vivo. These diagnostic methods could be potentially used for screening and staging of cancer as well as for monitoring of treatment. Formerly, mostly anatomical information played key role in medical visualization. Now, molecular visualization allows improving diagnostic parameters of standard diagnostic methods. Molecular imaging allows not only for localization of tumor, but also for visualization of biological processes that influence tumor behavior and response to therapy. This review reflects the potential role of radionuclide methods and radiopharmaceuticals in diagnostic and assessment of tumor response. The paper covers indications and capabilities of dedicated nuclear breast imaging systems such as breast-specific g-imaging and positron-emission mammography. The accuracy of different methods was analyzed. The analysis showed that new technological solutions allow to significantly increase informativeness of examinations through improved spatial resolution compared to whole-body imaging cameras. Molecular imaging is useful for neoadjuvant chemotherapy response monitoring and is highly sensitive for prediction of non-responsiveness during treatment of breast cancer. The sensitivity of molecular breast imaging is comparable with that of magnetic resonance imaging, but has higher specificity. Molecular imaging may play a great potential role in the diagnostic algorithm for breast cancer.

Известно, что дефицит гомологичной рекомбинации в опухолевых клетках, обусловленный, в основном, дефектом генов BRCA1/2, связан с высокой эффективностью лечения и благоприятным прогнозом заболевания. Однако наличие других альтернативных путей репарации ДНК, таких как активация генов NF-κB или PARP1, может оказывать дополнительный негативный эффект. Таким образом, целью работы явилась оценка связи аберраций числа копий ДНК генов BRCA1, NF-κB, PARP1 в опухолевой ткани молочной железы с эффектом химиотерапии и прогнозом заболевания. Материалы и методы. В исследование было включено 85 больных раком молочной железы IIA-IIIB стадии. ДНК выделяли из биопсийных образцов опухолевой ткани с использованием набора QIAamp DNA mini Kit (Qiagen, Germany). Было проведено микроматричное исследование всех образцов опухоли на ДНК-чипах высокой плотности фирмы Affymetrix CytoScanTM HD Array. Для оценки аберраций числа копий ДНК использовали программу «Chromosome Analysis Suite 3.3». Результаты. Было установлено, что наибольшая частота делеций наблюдается в гене BRCA1 (28%, 24 случая из 85), и это статистически значимо сопряжено с объективным ответом на неоадъювантную химиотерапии (p=0,02). Частота амплификации гена PARP1 в исследуемой группе больных составляет 62%, что также определяет хороший ответ на неоадъювантную химиотерапию вне зависимости от наличия аберраций других исследуемых генов. При анализе прогностической значимости аберраций генов было показано, что амплификация гена PARP1 является неблагоприятным маркером безметастатической выживаемости (log-rank test, p=0,02). Выводы. На основании полученных данных можно полагать, что аберрантное состояние генов BRCA1 и PARP1 в опухоли молочной железы, может также являться перспективным маркером эффективности химиотерапии и прогноза заболевания, что подтверждает актуальность исследования, но и требует дальнейшего детального изучения. It is well known that the presence in the tumor cells of such a phenomenon as a deficiency of homologous recombination, caused mainly by a defect in the BRCA1/2 genes, is associated with a favorable treatment effect and prognosis of the disease. But it has been established that the presence of other alternative ways of DNA repair, such as activation of the NF-κB or PARP1 genes, may have an additional negative effect. Thus, the aim of the work was to assess the association of chromosomal aberrations of the BRCA1, NF-κB, PARP1 genes in tumor breast tissue with the effect of chemotherapy and the prognosis of the disease. Materials and methods. The study included 85 patients with stage IIA - IIIB breast cancer. DNA was isolated from biopsy specimens of tumor tissue using the QIAamp DNA mini Kit (Qiagen, Germany). A microarray was studied for all tumor samples on Affymetrix CytoScanTM HD Array high-density DNA chips. To estimate the aberrations of the number of DNA copies, the program Chromosome Analysis Suite 3.3 was used. Results. It was found that the highest frequency of deletions is observed in the BRCA1 gene (28%, 24 cases out of 85), and this is statistically significantly associated with an objective response to neoadjuvant chemotherapy (p=0.02). The frequency of amplification of PARP1 in the studied group of patients is 62%, which also determines the presence of a good response to neoadjuvant chemotherapy, regardless of the presence of chromosomal aberrations of other have study genes. When analyzing the prognostic significance of gene aberrations, it was shown that PARP1 amplification is an unfavorable marker of metastatic-free survival (log-rank test, p=0.02). Conclusion. Based on the data obtained, it can be assumed that the aberrant state of the BRCA1 and PARP1 genes in a breast cancer may also be a promising marker of the effectiveness of chemotherapy and disease prognosis, which confirms the undoubted relevance of the study, but also requires further detailed study.

The purpose of the study was to analyze the 10-year outcomes after breast-conserving surgery with electronbeam intraoperative radiation therapy (IORT) followed by external beam radiotherapy (EBRT) in patients with breast cancer. Material and methods. The study included 905 patients with stage T1–2N0–1M0 breast cancer, who underwent breast-conserving surgery. Group I consisted of 746 patients who received IORT at a single dose of 10 Gy, biologically equivalent to 24.8 in standard fractionation, followed by EBRT at a total dose of 46 ± 8.1 Gy. A combined IORT and EBRT photon-equivalent dose delivered to the tumor bed was 60 Gy. Group II (control group) comprised 159 who received EBRT at a total dose of 40–44 Gy delivered to the remaining breast tissue and electron therapy delivered to the area of postoperative scar (a single 3–4 Gy fraction, 3 days per week, to the total photon-equivalent dose of 15-18 Gy). The combined photon-equivalent dose was 58 Gy. Results. According to RTOG/EORTC toxicity criteria, grade 1 radiation-induced damage was observed in 413 (55.3 %) patients of Group I and in 71 (44.6 %) patients of Group II (р=0.2); grade 2 radiation-induced damage in 76 (10.1%) and 43 (27 %) patients (p=0.0002) and grade 3 in 18 (2.4 %) and 9 (5.7 %) patients, respectively (р=0.1). Late radiation-induced injuries (grade I and 2) were mostly observed in the control group. Local recurrence was diagnosed in 11 (1.47 %) patients of Group I and in 14 (8.8 %) patients of Group II. The 10-year disease-free survival rates were 97.3 ± 1.08 % and 88.96 ± 2.8 % in Group I and Group II patients, respectively (р <0.05). The metastasis-free survival rates were 94.4 ± 2.7 and 90.46 ± 1.4 %, respectively (р<0.05). Conclusion. In patients with stage T1–2N0–1M0 breast cancer, combination of IORT and EBRT resulted in higher recurrence-free and metastasis-free survival rates than EBRT alone.

The study of functional polymorphism of the VEGF-A gene at positions –2578 in the promoter region and +936 in the 3`untranslated region in breast cancer patients with different receptor status of the tumor was undertaken to identify markers associated with risk of breast cancer. DNA samples from 292 women with breast cancer were analyzed. The genotyping of С–2578A and C+936T VEG-FA polymorphisms was performed using the method of restriction analysis of amplification products. The distributions of VEGF-A genotypes in patients having estrogen and progesterone receptors in tumor tissues were investigated and genotypes associated with risk of breast cancer recurrences were detected.

In the present study evaluated the distribution of leukocytes and cytotoxic T-lymphocytes in the stroma of various morphological structures of the invasive component of the breast cancer (IC NST) and association with tumor progression. Determined by immunohistochemistry CD3 +, CD45LCA + and CD8 + cells which are located near the tumor entities, as well as from those in the distance. Was determined relationship between presence of cytotoxic T-lymphocytes in the microenvironment of the discrete tumor cells, as well as in infiltrate the distance of any tumor structures and hematogenous, but not lymph node metastasis, and recurrence.

Background : Pathological complete response is a predictor of favorable clinical outcome in patients with tripl-negative breast cancer. Over the last years there was an increasing interest to the search for predictive markers of tumor response to neoadjuvant chemotherapy (NACH). The purpose of the study is to assess predictive markers: cytokeratin 5/6, epidermal growth factor receptor (EGF1) and proliferation marker Ki-67 in patients with basal-like tripl-negative breast cancer. Materials and methods : The study included 44 patients with basal-like tripl-negative breast cancer, who received 2–4 courses of NACH with FAC and CAX regimens. Estrogen and progesterone receptors, Her-2/neu, the levels of proliferative activity of Ki-67, CK 5/6 and EGFR1 were determined in all breast cancer biopsies. Response to NACH was assessed using RECIST scale. Results . Pathological complete response was observed in 69 % of breast cancer patients, who had only EGFR1expression in tumor tissue (р=0.01). The best response was noted at high proliferative activity of tumor cells (90 %, р=0.0006) and when using CAX regimen (81 %, р=0.004). The model of logistic regression enables prediction of pathological complete response with high rates of sensitivity (82 %) and specificity (67 %). Conclusion . The results obtained indicate that the studied markers can be used as predictors of clinical outcome and the devised model of logistic regression can be used to predict the chemotherapy response of breast cancer based on clinical pathological variables.

Hormone therapy with tamoxifen is the commonly used treatment for luminal breast cancer. However, it appears to be ineffective in 20–40 % of cases and the possible reasons of this failure are related to the features of distribution and structure of estrogen receptor alpha (ERα) in tumor tissue. Realization of the therapeutic effect of tamoxifen is carried out by blocking the activation center of AF-2 receptor. The change in the functional state of this receptor resulted from single-nucleotide polymorphisms coding 2228480 (G/A) in exon 8 of ERα gene is considered as a possible cause of treatment failure with tamoxifen. The purpose of the study was to analyze the relationship between the ERα expression and polymorphic variants in exon 8 of the ERα gene and the efficacy of tamoxifen in patientswith luminal breast cancer. Material and methods : The study included 97 patients with stage T1–2N0–1M0 luminal breast cancer, who received adjuvant chemotherapy with tamoxifen. The follow-up ranged from 24 to 130 months. Long-term treatment outcomes were assessed upon the progression of the disease with the evidence of distant metastases. In tumor tissue samples, the ERα expression was studied using the immunohistochemical method. The values of the ERα expression intensity as well as the character of ERα distribution were assessed. Polymorphic variants of exon 8 of the ERα gene were studied using real-time PCR. Results. The heterogeneous distribution of ERα gene was observed in 86.5 % cases with diseases progression and in 58.3 % of cases with favorable disease outcome (р=0.0072; χ2 =7.22). Mutation of rs2228480 (G/A) in exon 8 of the ERα gene was observed in 19.4 % of cases. Mutations were not noted in tumor cells with homogenous distribution of the ERα gene and mutations were found in 25.7 % (р=0.014; χ2 =6.09) in heterogeneous distribution. Mutation in exon 8 of the ERα gene was shown to occur more often in patients with disease progression (р=0.01; χ2 =6.52). Conclusion: The character of the ERα gene distribution and the presence of mutation in exon 8 of the ERα gene in tumor tissue can be considered as additional predictive factors of response to therapy with tamoxifen in patients with luminal breast cancer.

Background : Ethnic diversity of the population in the region of Siberia suggests the existence of different germline mutations in the BRCA1/2 genes associated with breast and ovarian cancer in different ethnic populations, but spectrum of these mutations has not been studied. Objective: Our aim was to evaluate the frequency of the most common mutations BRCA1 / 2 (BRCA1 5382insC, BRCA1 185delAG, BRCA1 4153delAG, BRCA1 T300G, BRCA2 6174delT) in women diagnosed with breast cancer among indigenous people and newcomers living in Siberia. Methods: We tested 1281 genomic DNA samples for the presence of BRCA1 5382insC mutation in patients diagnosed with breast cancer considering no family history. 72 patients having hereditary cancer signs were tested for the mutations BRCA1 185delAG, BRCA1 4153delAG, BRCA1 T300G, BRCA2 6174delT. Results: Out of 765 patients of Slavic ethnic group, 27 women (3.5%) were carriers of allele BRCA1 5382insC. The frequencies of mutations in patients with signs of hereditary cancer were: 8.3% in group of young patients (under 40 years), 20.0% in patients with bilateral cancer and 5.7% in patients with family history of breast or ovarian cancers. We tested 516 BC patients residing on the territory of the Buryat-Aginsky district, Republics of Tyva and Altai. Out of them, there were 197 patients among the indigenous population (buryats, tuvinians, altaians), and 319 patients among newcomers (Slavic ethnics). Mutations BRCA1 5382insC were detected only in women from Slavic ethnic groups. The frequency of BRCA1 5382insC mutation was 6% in the group where family history was excluded and 14% in the group of patients with characteristics of family cancer. Allele BRCA1 5382insC was not found in indigenous breast cancer patients, although 59 patients had signs of hereditary cancer. In women from Slavic ethnic group, the BRCA1 185delAG, BRCA1 4153delAG and BRCA1 T300G mutations were detected in 9.1% of cases and were not found in patients among the indigenous population. Conclusion: studies of mutations in the BRCA1 gene in breast cancer patients from Siberia confirmed data on the high frequency of «founder mutation» BRCA1 5382insC in Slavic population and indicate the advisability of further studies to identify the genes responsible for the occurrence of hereditary breast cancer in the indigenous population.

For the first time in a comparative perspective the epigenetic status of the benign proliferative processes, breast cancer, and metastases to regional lymph nodes was studied using DNA methylation microarray "GoldenGate Cancer Panel I" ("Illumina", USA). The functional groups of differentially methylated genes were identified in each set of samples. The genes that regulate cell proliferation and mobility were methylated in samples with benign proliferative processes. An aberrant methylation of the genes responsible for cell differentiation and proliferation, as well as protein phosphorylation and cell mobility was observed in the samples with malignant phenotype. Differential methylation of the genes that regulate cell adhesion, the formation of anatomical structures, angiogenesis, immune response, signal transduction, and protein phosphorylation was found in the samples with metastases to regional lymph nodes in comparison with the morphologically unaltered breast epithelium. The tissues from the benign proliferative processes and metastases to regional lymph nodes were generally characterized by a relatively lower level of epigenetic variability in comparison with the tissues of the primary tumor.

Experimental and clinical evidence suggests that the immune system when exposed to conventional cancer chemotherapy is involved in the antitumor effect. A study is conducted to assess the relationship between immunological parameters and effectiveness of neoadjuvant chemotherapy (NAC) in breast cancer (BC) patients. The study included 269 patients with BC (T1–4N0–3M0) and 24 practically healthy comparable age women. The estimation of the subpopulation composition of blood mononuclear cells, their functional activity, apoptosis markers, allelic polymorphism of cytokine genes, depending on the presence or absence of clinical response to NAC was done. Complete tumor regression was associated with an increase in the number of cytotoxic CD8+-cells, high functional activity of lymphocytes (proliferation in response to mitogen, the secretion of cytokines TNFα, IL-1β and IL-10, IFN-γ) and neutrophils. Close relation of highly functional cytokine genotype and high cytokine secretion in blood cells with an objective clinical response to chemotherapy was revealed. Thus, the findings suggest that an objective clinical response to NAC is associated with structural and functional preservation of the immune system. Constitutive characteristics of the patient’s organism, responsible for the level of expression of pathogenetically relevant cytokines that play a key role in the functioning of the immune system are have important meaning.

Previously, we showed the association of neoadjuvant chemotherapy (NAC) response with changing the expression vector (increase or decrease) of multidrug resistance genes (MDR) in breast tumors during chemotherapy. The aim of the present study was to evaluate the relation between changes in the expression vector of MDR genes and distant metastasis-free survival. Patients (n = 120) with breast cancer (T1-4N0-3M0) treated by 2-4 cycles of NAC (CAX, FAC, and taxane regimes) and 4 cycles of adjuvant chemotherapy (FAC) were included. TaqMan-based quantitative reverse transcriptase PCR (qRT-PCR) was used to estimate the expression of the following MDR genes: ABCB1, ABCC1, ABCC2, ABCC5, ABCG1, ABCG2, GSTP1, and MVP--in biopsies before NAC and in tumor samples after chemotherapy. Comparing the corresponding expression levels allowed us to identify the vector of expression change during NAC. The results showed that 5-year distant metastasis-free survival was 73-78% in patients with a decrease in ABCB1, ABCC2, and ABCG1 expression. The up-regulation of these genes during NAC was related to a significant decrease (up to 50-55%) in metastasis-free survival (Kaplan-Meier analysis: log-rank p value = 0.006-0.03). The association of changing the expression vector of MDR genes with metastasis-free survival did not depend on tumor size, lymph node involvement, histological form, receptor status, molecular subtype, and others clinicopathological parameters of breast cancer. The obtained data suggest that changing the expression vector of MDR genes in breast tumors during NAC may be used as a new potential prognostic marker of breast cancer. An increase in tumor expression of ABCB1, ABCC2, and ABCG1 during chemotherapy is a factor of poor prognosis, whereas down-regulation of these genes--a favorable prognostic marker.

Introduction. 199Tl-chloride scintigraphy is used for visualization of malignant tumors and benign lesions including inflammation. Differentiating of them is performed by permanent and independent retention index, but not always effective. Purpose: improve the differentiating of malignant tumors from benign lesions using 199Tl-chloride scintigraphy. Materials and methods. Results of 97 patients’ 199Tl-chloride scintigraphy with 119 regions of hyperfixation in malignant tumors (n=68) and benign lesions (n=51) were studied. Lesions were localized in musculoskeletal system (n=89), in lungs and mediastinum (n=25) and other organs (n=5). Results. Sensitivity, specificity, positive, negative predictive value and accuracy 199Tl-chloride scintigraphy in differentiating of malignancies from benign lesions were 75.0%, 86.3%, 58.1%, 93.2% and 84.0%, respectively using optimal value of permanent and independent retention index RI=0.89 (RI=DR/ER). Significant correlations between retention index (RI) and 199Tl-chloride hyperfixation in early phase of scintigraphy (ER) in group with malignancies (r=-0.70, p=0.00001) and benign lesions (r=-0.76, p=0.00001) were revealed. As a consequence, method for differentiating malignant tumors from benign lesions with optimal dependent relative retention index RI(relative)=RI+0.055-ER and cut-off value RI(relative)=0.98 4 was proposed (sensitivity, specificity, positive, negative predictive value and accuracy 199Tl-chloride scintigraphy in differentiating of malignancies from benign lesions were 83.8%, 86.3%, 73.8%, 92.0% and 85.5%, respectively). Statistically significant (p=0.01) improvement of efficiency of 199Tl-chloride scintigraphy diagnostics in differentiating of malignant tumors from benign lesions using relative retention index (RI(relative)) compared with independent retention index (RI) were revealed by ROC-analysis. Conclusion. The optimal value of permanent and independent retention index for differentiating of malignant tumors from benign lesions are RI=0.89. Efficiency of 199Tl-chloride scintigraphy diagnostic in differentiating of these processes improves using of relative retention index.