Лабораторные и экспериментальные исследования. Рубрика в журнале - Сибирский онкологический журнал
Loss of heterozygosity in the BRCA1 and BRCA2 locus in breast cancer
Статья научная
One of the factors of variability of malignant neoplasms is the loss of heterozygosity (LOH). The biological meaning of LOH, in relation to carcinogenesis, is associated with the inactivation of heterozygous loci of pathogenetically significant genes. Thus, the aim of this work was to study BRCA1/2 LOH in breast tumors. Material and Methods. The study included 122 patients with stage IIAIIIC breast cancer. DNA was isolated from 122 biopsy samples of tumor tissue using the QIAamp DNA mini Kit (Qiagen, Germany). To assess the status of LOH, microarray analysis was performed on high-density DNA chips from Affymetrix CytoScanTM HD Array. To process the results of microchipping, we used the Chromosome Analysis Suite 3.3 program (Affymetrix, USA). Results. The loss of heterozygosity in the BRCA1 gene was found to be associated with response to NAC. It was shown that in 59 patients LOH in the BRCA1gene was associated with an objective response to treatment (p=0.005). The presence of LOH in the studied genes was associated with a favorable prognosis. The 5-year non-metastatic survival rates were 75 % and 100 % in patients with LOH in the BRCA1 and BRCA2 genes, respectively (log-rank test: p=0.003 and p=0.05, respectively). Conclusion. The phenomenon of LOH in the BRCA1/2 genes was shown to be associated with response to NACT. BRCA1/2. Further studies are needed to evaluate the frequency of BRCA1/2 LOH after NAC for choosing and changing treatment tactics.
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MIR-155 и MIR-223 как маркеры клинических особенностей течения хронического лимфолейкоза
Статья научная
Введение. Хронический лимфолейкоз (ХЛЛ) - заболевание, которое характеризуется высокой межиндивидуальной гетерогенностью как клинического течения, так и молекулярных паттернов экспрессии генов и регуляторных РНК, способных оказывать влияние на патологический процесс. Вовлеченность регуляторных miR-155 и miR-223 в патогенез ХЛЛ в целом известна, но информации о возможных колебаниях экспрессии miR-155 и miR-223 в зависимости от динамики развития патологического процесса и характеристик врачебного вмешательства недостаточно. Цель исследования - изучение уровня экспрессии miR-155 и miR-223 у больных ХЛЛ с разными биологическими, клиническими особенностями и характеристиками проводимой терапии в субстратах периферической крови: плазме, лимфоцитах, внеклеточных везикулах, а также в костном мозге. Материал и методы. При выполнении работы использовались образцы периферической крови и костного мозга 38 пациентов с диагнозом ХЛЛ из Городского гематологического центра ГБУЗ Новосибирской области «Городская клиническая больница № 2» с 2016 по 2017 г. Оценка уровня экспрессии miR-155 и miR-223 проводилась методом ОТ-ПЦР в реальном времени по принципу TaqMan. Оценку значимости различий групп проводили с помощью непараметрического критерия Манна-Уитни либо непараметрического критерия Краскела-Уоллиса с последующим попарным сравнением с помощью критерия Манна-Уитни.
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Статья научная
Activating mutations of egfr are associated with sensitivity of non-small cell lung cancer (NSCLC) to tyrosine kinase inhibitors (TKI). Liquid biopsy using circulating cell-free tumor DNA (cfDNA) is proposed in cases when formalin fixed paraffin embedded (FFPE) tumor tissue is not available and for monitoring of egfr status. In the study we evaluated new qPCR assay for egfr mutations in plasma cfDNA. Sensitivity of the assay was 1 % of the mutant allele for L858R, L861Q, S768I mutations and deletions in exon 19, and 5 % of the mutant allele for G719X or T790M mutations Before surgery, mutation was detected in plasma of 4 out of 7 patients (57 %) with mutant egfr in FFPE tumor tissue. Mutations found in cfDNA completely matched those found in tumor tissue in 2 cases. In one case with G719X and S768I mutations in FFPE tissue, only S768I was found in cfDNA. In another case, T790M was detected in plasma in addition to L858R that was present in tumor tissue. No egfr mutations were detected in plasma DNA from 12 healthy volunteers and 13 cases of NSCLC with wt egfr suggesting 100 % specificity of the assay. Liquid biopsy detected egfr mutations in cfDNA in 8 of 16 cases of NSCLC with mutant egfr being under therapy with TKI. Among them, 7 cases had mutations in liquid biopsy that matched those in tumor tissue and another case had T790M in addition to L858R. In 3 cases increased mutant allele frequency was detected 212 months before clinical progression.
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Morphological heterogeneity of intratumoral macrophages in prostate tumors
Статья научная
Background. Prostate cancer (PCa) is the most common human cancer worldwide. in the progression of prostate cancer, the total number of macrophages in the tumor tissue is associated with poor prognosis and increased risk of metastasis. However, the heterogeneity of intratumoral macrophages at various stages of PCa development, and the role of tumor-associated macrophages (TAMs) have been insufficiently investigated. The aim of the study was to analyze the morphological features, size and number of TAMs in PCa tissue samples, and to reveal their correlation with clinical data of patients. Material and Methods. immunohistochemical analysis of 36 paraffin blocks of patients with PCa (pT2a-3bN0-1M0) was performed using antibodies to the scavenger receptor CD68. Results. Foamy CD68+ macrophages were found in the tumor tissue. The indicator "number of macrophages per total number of fields of view with macrophages" was the lowest in patients with a Gleason score of 6 (5.8) (11.0 - in patients with a Gleason score > 8). Macrophages formed larger clusters in patients with severe PCa. Small but not large macrophages were significantly more common in patients with lymph node metastases (48 vs 24 in the N0 group; p=0.14). The number of small macrophages (smaller than 100 pm2) increased in a series of patients with Gleason scores of 6, 7 and > 8 (24, 47.5, 72, respectively, p=0.052). Conclusion. As the tumor process progressed and the risk of biochemical recurrence increased, there was a trend towards an increase in the total area of large, foamy TAMs, presumably rich in lipids, as well as wider distribution of small macrophages with a tendency to form clusters. We hypothesize that foamy macrophages are involved in the further recruitment of small TAMs, subsequently leading to metastasis and tumor progression.
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Статья научная
Introduction. In the development of new methods of radiotherapy, studies of the biological effects of sparsely (photons, electrons) and densely (protons, ions) ionizing radiation are relevant. Reproducibility is a challenge in preclinical studies. Dosimetric phantoms of laboratory animals are an effective tool for dose assessment, facilitating standardization of tests conducted under different conditions. Existing phantoms often fail to address radiobiological issues like placing of biological samples or dosimetry detectors. A method for manufacturing dosimetric phantoms must be developed to accurately manufacturing products and modify their design in accordance with the task.
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Статья научная
The mechanism of the relationship between pretumor changes in the bronchial respiratory epithelium and the risk of progression of non-small cell lung cancer (NSCLC) remains unclear. It has been suggested that the relationship between reactive changes in the bronchial mucosa and NSCLC progression may be caused by the functional status of monocytic-macrophage cells as important participants in inflammation, which determines both the risk of premalignant changes in the epithelium and malignant progression. The purpose of the study was to investigate the phenotypic profile of peripheral blood monocytes and macrophages induced from monocytes in vitro depending on the state of respiratory epithelium in NSCLC patients. Material and Methods. The study included 39 patients with newly diagnosed NSCLC. Based on the morphological examination of small bronchi taken at the distance of 3-5 cm from the tumor, patients were divided into the following groups depending on the type of pretumor changes: no pretumor changes (n=6), isolated basal cell hyperplasia (BCH) (n=13), combination of BCH and squamous metaplasia (SM) (n=3), combination of unchanged epithelium and focal BCH (n=17). The phenotypic features of peripheral blood monocytes and in vitro-induced macrophages were assessed before treatment using flow cytometry. Results. The state of the respiratory epithelium in NSCLC patients prior to the start of anticancer treatment was associated with the phenotypic features of peripheral blood monocytes, but not with the profile of macrophages induced from them. Distortion of the response of induced macrophages to the polarizing stimuli was observed in NSCLC patients: the cultured cells responded to both M1 and M2 inducers (LPS and IL-4, respectively) with a phenotype shift to M2, while the CD206 marker expression varied depending on the presence and type of pretumor changes. Conclusion. The phenotypic profile of peripheral blood monocytes was associated with the state of the respiratory epithelium in NSCLC patients before anti-tumor treatment, but not with the phenotypic features of induced macrophages.
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Статья научная
The aim of this study was to investigate the potential of compounds from the breadfruit tree (Artocarpus altilis) as anti-breast cancer agents using in silico techniques. Material and Methods. The methods used in this study include molecular docking and ADMET (Absorption, Distribution, Metabolism, Excretion, and Toxicity) predictions to evaluate the interactions with Estrogen Receptor alpha (ERα). Results. Among the 22 compounds tested, Cycloaltisin-7 exhibited the most favorable binding affinity, with a free energy of -10.25 kcal/ mol and an inhibition constant of 30.89 nM. Additionally, Cyclocommunol and Cudraflavone B demonstrated significant binding interactions, with free energies of -9.61 kcal/mol and -9.53 kcal/mol, and inhibition constants of 90.82 nM and 103.50 nM, respectively. For comparison, the standard compound 4-Hydroxytamoxifen showed superior binding characteristics, with a free energy of -12.36 kcal/mol and an inhibition constant of 867.83 pM. ADMET predictions indicate that Cycloaltisin-7, Cyclocommunol, and Cudraflavone B meet essential drug-like criteria, suggesting their potential as viable candidates for further development as breast cancer therapeutics. Conclusion. These findings highlight Cycloaltisin-7 as a particularly promising compound, with Cyclocommunol and Cudraflavone B also showing considerable potential. This research provides valuable insights for the advancement of plant-based treatments for breast cancer. Supporting agencies: This study was funded by APBU 2024 Research Grant Universitas Islam Kalimantan MAB Banjarmasin, Indonesia.
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Primary liver cancer in the Republic of Sakha (Yakutia)
Статья научная
Territorial and ethnic features of the prevalence of primary liver cancer in the republic of Sakha have been studied. The republic has been shown to be a high-risk area for primary liver cancer, which is the 2-nd most common form of cancer in males and the 3-rd most common form of cancer in females among all digestive system malignancies. The standard liver cancer incidence rates are 5 times higher than the average incidence rates in Russia. Primary liver cancer is more oftern registered in the age group of 70 years and more. Incidence rates are mainly raised due to indigenous and rural populations. The liver cancer incidence rates show a tendency to stabilization all over the republic.
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Статья научная
PCR clamping/wild-type blocking PCR with non-extendable locked nucleic acid (LNA) oligonucleotides is used for sensitive detection of somatic mutations in tumors. Various versions of the technique use different DNA polymerases and LNA oligonucleotides with and without additional phosphorothioate modifications. Here we studied requirements for successful PCR clamping with LNA oligonucleotides and Taq DNA polymerase for analysis of mutations in KRAS and bRAF genes by means of real-time PCR and Sanger sequencing. We found that addition of phosphorothioate linkages at the 5’-end of LNA oligonucleotide to protect from 5’-exonuclease activity of Taq DNA polymerase did not improve clamping. For most target sequences, efficient clamping was observed at melting temperature of LNA oligonucleotide 20-25°C above annealing/extension temperature of the PCR with a 2-step protocol. Under such conditions, simple and sensitive detection of mutations in KRAS and bRAF genes was feasible using real-time PCR with TaqMan probes or Sanger sequencing.
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Study of dendritic cell migration using CELL-IQ analysis system
Статья научная
Dendritic cells (DCs) belong to specialized pool of antigen-presenting cells with high functional plasticity and manifest with immunostimulatory or immunosuppressive potential depending on sequence and combination of microenvironment stimuli, which determine their differentiation, maturation and activation. The use of antitumor DCs vaccines is based on the ability of DCs specifically activated in vitro migrate for antigen presentation to T-lymphocytes. We studied the components of the tumor microenvironment that are capable of inhibiting DCs migration. The study of the mobility of DCs in Cell-IQ experimental analytical system showed the presence of an inverse correlation of high strength between the average trajectory speed and the level of immunosuppressive factors (ISFs) in supernatants of cultured skin melanoma cells (TgFβ1, IL-10, IL-18, VEgF-A, EgF, FgF, HgF, sFASL (p
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Study of the effect of lomustin on HER2-positive breast cancer in FVB/N her-2 transgenic mice
Статья научная
Because of the high risk of brain metastases from HER2-positive breast cancer, the study of the anticancer activity of drugs used to treat brain tumors, in particular lomustine, is of great importance. In the FVB/N Her-2 transgenic mice bearing HER2-positive breast cancer (BC HER2+), a single oral administration of lomustine at a dose of 50 mg/kg resulted in a significant tumor growth inhibition (up to 96 %, p
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Статья научная
Introduction. Identification of persons with a high oncological risk to squamous cell carcinoma of the head and neck region is an urgent problem for the early diagnosis of this disease. The activity of circulating proteasomes can be a criterion for predicting the risk of the larynx and oral cavity cancers in patients with precancerous diseases of the upper respiratory and gastrointestinal tracts. The aim of the study is to investigate the chymotrypsin-like and caspase-like activities of circulating serum proteasomes depending on the localization of precancerous and neoplastic diseases of the larynx and oral cavity. Material and Methods. The study population consisted of 35 patients with histologically verified HNSCC (T1-3N0-3M0), 15 patients with chronic hyperplastic laryngitis (CHL) and oral leukoplakia, and 10 healthy volunteers who did not have chronic upper respiratory tract diseases in the acute stage. The median age of the patients was 53 ± 5.3 years. Results. An increase in the studied proteasome activities was found in the blood serum of patients with malignant tumors as compared with patients with chronic hyperplastic diseases associated with precancerous changes, as well as in the larynx and oral cavity cancers groups as compared with healthy donors. At the same time, depending on the localization of the pathological process, it was shown that only the chymotrypsin-like activity of the circulating pool of proteasomes significantly differs both in the groups of oral cancer leukoplakia, and in the groups of laryngeal cancer chronic hyperplastic laryngitis with dysplastic epithelial lesions. In addition, differences were found between chymotrypsin-like and caspase-like activities of circulating serum proteasomes in patients with chronic hyperplastic laryngitis with oral dysplasia and leukoplakia. Conclusion. The results obtained indicate that the determination of the CTP activity of the circulating pool of proteasomes can be used as a criterion for predicting the risk of the larynx and oral cavity cancers in patients with precancerous diseases of the larynx and oral cavity.
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Статья научная
Localized and metastatic tumors are known to lead to the formation of circulating tumor cell (CTC) clusters in the blood. Currently, there is a heightened interest in the study of molecular and biological characteristics of CTCs. Recent studies have shown the presence of different populations of CTCs in the blood of cancer patients. Some cells are cancer stem cells, some tumor cells undergo epithelial-mesenchymal transition (EMT), and most CTCs do not have features of either stem cells or EMT. The aim of the study was to evaluate the five-year metastasis-free survival rate in patients with invasive breast carcinoma, depending on the presence of various populations of circulating tumor cells in the blood before treatment. Material and Methods. A prospective study included 47 patients with newly diagnosed invasive breast cancer (T1-4N0-3M0), who were treated at Cancer Research institute, Tomsk National Research Medical Center. The patients aged 31 to 69 years. The presence of different populations of CTCs in the blood of patients before treatment was determined by multicolor flow cytometry on the BD FACS Canto system, using different fluorochrome-labeled monoclonal antibodies to EpCam, CD45, CD44, CD24, and N-cadherin. Five-year metastasis-free survival was evaluated by the Kaplan-Meier method. The differences were considered significant at p function show_abstract() { $('#abstract1').hide(); $('#abstract2').show(); $('#abstract_expand').hide(); }
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The role of EZH2 and ARID1A in the diagnosis of flat urothelial lesions with atypia
Статья научная
Background. Diagnosis of urothelial carcinoma in situ is of great importance because it has prognostic and therapeutic value. We aim to determine the utility of EZH2 and ARiD1A as a new tool in the diagnosis of carcinoma in situ. Material and Methods. This retrospective cross-sectional study included Twenty-four specimens of flat urothelial lesions, twenty specimens of CiS, and 10 of normal adjacent urothelium that was taken by cystoscopic resection biopsy procedure. immunohistochemical expression of EZH2 and ARiD1A. were evaluated in all studied cases. Results. All normal urothelium specimens showed high nuclear staining for ARiD1A and negative nuclear staining for EZH2. High EZH2 expression was observed in 80 % of CiS specimens compared to 20 % of flat urothelial lesions with atypia (p=0.001 ), while high ARiD1A expression was observed in 70.8 % of flat urothelial lesions with atypia compared to 25 % of CiS specimens (p=0.001). EZH2 was more accurate and specific in the diagnosis of carcinoma in situ. Conclusion. EZH2 and ARiD1A are promising diagnostic markers for urothelial CiS. EZH2 is more accurate and specific than ARiD1A in the diagnosis of carcinoma in situ versus other flat urothelial lesions.
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Статья научная
Hereditary breast cancer (HBC) is a heterogeneous disease caused by mutations in genes characterized by ethnic specificity. The clinical heterogeneity of this disease significantly complicates its diagnosis. The use of high-throughput sequencing is one of the approaches that allow the search for genes and their variants associated with the development of HBC. The purpose of the study was to search for new genes associated with HBC in the understudied ethnic groups of Siberia by using whole exome sequencing (WES). Material and Methods. WES was performed on a cohort of 16 probands with BC (Tuvan, Yakut, Altai ethnos). The study material was genomic DNA isolated from peripheral blood leukocytes. Libraries were prepared using a BGI Optimal DNA Library Prep kit. An Agilent SureSelect Human All Exon V6 kit was used for hybridization. High-throughput sequencing was performed on a DNA nanoball sequencing platform (DNBSeq-G400).
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Статья научная
Аберрантное метилирование промоторных участков генов - основное эпигенетическое изменение, характеризующее колоректальные неопластические образования. В настоящей работе был исследован количественный уровень метилирования 42 CpG-сайтов промоторных участков генов MGMT, APC и CDH13 в опухолях толстой кишки по отношению к уровню метилирования прилежащей условно нормальной ткани 25 пациентов. С помощью метода пиросеквенирования выявлено повышение уровня метилирования промоторных участков генов MGMT, APC и CDH13 в опухолевых образцах от 3 до 5 раз. В этих же образцах опухолей проведен скрининг активирующих SNP-мутаций в онкогенах KRAS (40 %), NRAS (0 %) и BRAF (0 %). Наличие SNP-мутаций в гене KRAS сопровождалось гиперметилированием одного или более промоторов исследованных генов. Доказана ассоциация этого эпигенетического показателя с метастазированием опухоли. Полученные данные об увеличении метилирования промоторных участков генов-онкосупрессоров могут быть использованы в качестве чувствительных прогностических маркеров прогрессирования и метастазирования колоректального рака.
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Статья научная
Найдены параметры линейно-квадратичной модели для нейтронной терапии. На этой основе проведено сравнение значений однократных терапевтических доз в нейтронной терапии, рассчитанных по модели время - доза - фракционирование и по линейноквадратичной модели. Установлено, что результаты расчета хорошо согласуются между собой. Также проведено сравнение степени расхождения результатов расчета по модели время - доза - фракционирование и по линейно-квадратичной модели в нейтронной и гамма-терапии. Показано, что применение линейно-квадратичной модели, разработанной для нейтронной терапии, способно обеспечить адекватный контроль за лучевыми реакциями облучаемых тканей.
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Статья научная
Цель. Изучение уровня адипокинов сыворотки крови, экспрессии AdipoR1, AdipoR2 и sOb-R у больных раком ободочной кишки в зависимости от клинико-морфологических параметров и наличия метаболического синдрома (МС). Материал и методы. В исследование включен 31 больной раком ободочной кишки I-III стадий с МС и без метаболических нарушений. Сывороточный уровень лептина, адипонектина, AdipoR1, AdipoR2 и растворимой формы рецептора лептина оценивался с помощью ELISA. Результаты и заключение. Частота МС у больных раком ободочной кишки составила 71 %. Выявлены взаимосвязи уровня лептина сыворотки крови с полом и стадией заболевания. Показана тенденция к повышению лептина у больных с МС. Уровень sOb-R был выше у больных без МС. Уровень AdipoR1 и AdipoR2 был значимо выше у больных раком ободочной кишки женского пола. Имелась тенденция к повышению рецепторов адипонектина у больных раком ободочной кишки с наличием МС.
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Активация сигнального пути интерферона-альфа ресвератролом, генистеином и кверцетином
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Ресвератрол, кверцетин и генистеин, относящиеся к полифенолам вторичных метаболитов растений, обладают антиканцерогенным и противовирусным эффектами, реализуемыми в результате их плейо-тропного действия на различные макромолекулы клетки. Эти соединения могут взаимодействовать с ДНК, не образуя ковалентные связи. При этом может происходить изменение пространственных, физико-химических и структурных характеристик ДНК, что может приводить к нарушению функциони-рования белков метаболизма ДНК и вызывать дестабилизацию хроматина. Такие эффекты были опи-саны для нового противоопухолевого препарата Кураксина CBL0137, причем индуцированная данным соединением дестабилизация хроматина приводила к активации сигнального пути интерферона-α. Используя клеточную линию HeLa с трансгенным флуоресцентным белком mCherry, содержащим в промоторной области консенсусный сайт связывания интерферона-α (ISRE), мы продемонстрировали дозозависимый стимулирующий эффект ресвератрола, кверцетина и генистеина на активность сигналь-ного пути интерферона-α. Использование прижизненной флуоресцентной микроскопии на клеточной линии HT1080 c трансгенным флуоресцентно-меченным гистоном H1.5 позволило продемонстриро-вать, что данные полифенолы вызывают перераспределение данного линкерного гистона в ядрах клеток. Полученные нами данные свидетельствуют о возможности существования ДНК-зависимого механизма реализации противоопухолевого действия растительных полифенолов и необходимости дальнейшего изучения влияния полифенолов на структуру хроматина и связанного с этим изменения функционирования генома, в частности регуляции сигнального пути интерферона-α.
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Цель исследования - сравнить показатели функциональной активности натуральных клеток-киллеров в зависимости от наличия злокачественного процесса и его распространенности. Материал и методы. В исследование были включены 10 больных раком яичников IIIC стадии, 5 больных с доброкачественными опухолями (ДО) яичника, 15 больных колоректальным раком TNMстадии. Контрольную группу 2-40-20 составили 5 обследованных здоровых доноров. Для оценки количества и функциональной активности NK-клеток периферической крови и асцитической жидкости (АЖ) выполнена многоцветная проточная цитометрия на цитофлуориметре FACS Canto II. результаты. Наличие злокачественного новообразования сопровождалось статистически значимым снижением в периферической крови относительного количества активированных NK-клеток, секретирующих гранзим В (GB) (CD56+CD107a+GB+PF-), и увеличением доли дегранулированных NK-клеток (CD56+CD107a+GB-PF-) по сравнению с соответствующими показателями у здоровых лиц. При этом отличительной особенностью больных раком яичников были низкие показатели общего количества NK-клеток в периферической крови (p
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