Biobanks are the platform for innovative biomedical research in the field of translational and personalized medicine. The important aspect for conducting large-scale research in the field of genomics, transcriptomics, proteomics is the availability of a sample information of the documented high-resolution samples. The biobanks solve the problem of forming groups of patients with different nosology within the population of interest and provide clinical and laboratory information on each sample. The aim of this article is to describe the Biobank processes of the National Medical Research Centre for Oncology within the framework of existing projects and build up collections. The review discusses the main stages of the systematized biobank process, describes the methods of sample preparation of different types of biological material, and also provides statistics of the build up collections. To this date, the predominant part of the depository consists of tissue samples of patients diagnosed with colorectal cancer 24% and stomach cancer 23% of the total number of tissue samples, while the number of tissue samples of pancreatic cancer is 10%, and esophageal cancer and breast cancer 22%. In addition to tissue samples, the biobank of the National Medical Research Centre for Oncology stores 24 cell lines of a human origin and the collection of 200 microbiota samples: 100 are from patients diagnosed with lung cancer and 100 from conditional healthy donors. Currently, the studies have been performed on biomaterial from the biobank build up collections to search for prognostic biomarkers and potential targets for targeted therapy by using high-throughput sequencing in patients diagnosed with pancreatic cancer and brain cancer. Thus, the collections play an important role for research in the field of personalized medicine, providing early diagnosis and effective treatment for each patient.
Experimental dependency of the photosystem's response on the wavelength of exciting radiation, also known as action spectrum, may be substantially affected by the spectrum shape of this radiation. This is especially important in the case, when different radiation sources are used for the investigation of action spectrum. For instance, too wide emission peaks of radiation sources can blur the scopes of actual action spectrum and distort information about the properties of photosystem at certain wavelength regions. Here, we propose a method for the correction of experimental action spectrum by the recalculation of experimental data of photoresponse according to actual spectra of exciting radiation. In the case of overlapping radiation spectra from different radiation sources, this method results in much better correlation of experimental action spectrum to actual action spectrum or absorption spectrum of photosystem. The data on photoactivity of several photocatalysts are presented to illustrate and validate the proposed method. Activity of photosystem depends on the actual spectrum of the radiation source Single-peak optical radiation sources with the same basic wavelength may cause a different photoactivity Effect of actual spectrum of the light source on the photoactivity is to be considered
e15045 Background: Berberine is an alkaloid compound with a structure that is highly similar to that of intercalating agents. It affects numerous cell signaling pathways and is widely studied as potential anticancer drug. It is known that berberine affects cancer cells migration through metalloproteinase-2 inhibition, but this effect was never studied on glioma cells. Anti-migratory drugs are of special interest in brain cancer therapy since glioma's highly invasive nature makes total surgical removal of tumor practically impossible. The aim of the study was to evaluate berberine anti-migratory activity on glioma cells. Methods: Cell migration capacity of T98G and U87MG cell lines, as well as primary glioma cell culture established in our laboratory, was assessed via standard wound healing assay with automated image acquisition and analysis on Lionheart FX (BioTek) cell imager. Prior to assay setting up cell cultures were maintained in DMEM medium with L-glutamine (1 μM) (Gibco) and 10% FBS (Gibco) at 37C0 and 5.0% CO2. Cells were seeded at 250 000 cells per well on 24-well plates and incubated overnight in order to attach to plate bottom. After that a vertical wound was made manually in each well, and berberine was added to experimental wells to final concentration 50 mg/L. Plates with cells were continuously incubated and photographed in cell imager at 37C0 and 5.0% CO2. The extent of cells migration was measured as the percent of wound area decrease after 24 hours of incubation in relation to starting time point. Data are given as: Mean ± 95% confidence interval. Results: In our study we berberine exhibited anti-migratory activity in all cell cultures under study. In rather fast growing primary cell culture wound area decrease was 99.23%±0.62% in control sample and 91.75%±0.28% in experimental sample. The difference was small but significant at p < 0.001 level (df = 30). Popular permanent glioma cell lines T98G and U87MG showed more prominent decrease in studied parameter with higher degree of variance at the same time. In T98G wound area decrease was 71.6%±12.3% in control and 48.8%± 7.6% in experimental samples after 24 hours of cultivation in presence of 50 mg/L berberine. While U87MG demonstrated 60.28%±5.13% and 37.5%± 8.34% wound area decrease accordingly. The obtained difference between control and experimental groups in permanent cell cultures was statistically significant at the 0.05 level (df = 30). Conclusions: Our preliminary research proved berberine to be potent anti-migratory agent in glioma treatment. Further investigations are needed to evaluate its ability to inhibit glioma cell expansion in vivo.
The Biobank of the National Medical Research Center of Oncology is a multi-layered infrastructure with large collections of biological samples, complemented by extensive and well-annotated clinical and pathological patient data, including medical images, pathological histology, and molecular analysis of biosamples. To date, the biobank of the National Medical Research Center of Oncology contains collections of primary and immortalized cancer cell lines of human origin. The collection of primary cell lines was formed from samples of postoperative material taken during the removal of tumors of various localizations (breast cancer, prostate cancer, lung cancer). All cell lines underwent internal quality control for contaminants (exogenous viruses, mycoplasmas and bacterial L-forms), viability and were cultivated without antibiotics. On the basis of the collected samples, a significant number of projects in the field of biomedicine were carried out, the results of which are described in this article.
e15009 Background: Multiple myeloma (MM) is a B-cell malignancy resulting from the abnormal proliferation of neoplastic plasma cells that produce monoclonal immunoglobulin (Ig). The high variability of the course of this disease, its genetic clonal heterogeneity, is due to chromosomal deletions, chromosomal hyperploidy involving an odd number of chromosomes, as well as genetic aberrations, such as rearrangement of the Ig heavy chain gene loci. Since the available biomarkers do not take into account this feature of MM, there is a need to develop more advanced biomarkers that will more accurately predict the course of the disease and response to treatment. Methods: The collection of MM samples included biological samples obtained from patients over 18 years of age with a diagnosis of MM, who received treatment in the National Medical Research Center of Oncology since 2019. Only patients who signed an informed consent for the use of their biomaterial for scientific purposes were included in the project. The material was collected according to the developed algorithm of clinical information and biological material collection, sample preparation, quality control and storage in the cryostorage of the National Medical Research Centre for Oncology of the Ministry of Health of Russia (Rostov-on-Don). Results: As of December 23, 2021, collection consists of 387 samples of whole blood, serum, plasma and mononuclear cells obtained from 42 MM patients of both sexes, whose average age was 59.7 ± 1.49 (±SD) years. Each patient was assigned a unique identification number. Freezing of the obtained samples occurred in accordance with the low-temperature storage protocol. Registration, accounting and certification of the material were carried out in a specialized database for recording and storing information about biological samples. Conclusions: The identification of MM biomarkers is important for increasing the sensitivity of molecular monitoring, which makes it possible to stratify patients into risk groups for early relapses and treatment resistance development. Thanks to the accumulated experience, the Biobank of the National Medical Research Centre for Oncology of the Ministry of Health of Russia serves as a valuable resource for providing research in the development of new predictive molecular markers.
Цель исследования. Исследовать влияние покрытия из СИЭЛ 159-330 на скорость и характер образования клеточных скоплений в висячей капле в сочетании с применением метилцеллюлозы (МЦ) и коллагена в качестве агентов, улучшающих агрегацию клеток. Материалы и методы. Клетки культуры рака молочной железы ВТ20 в количестве 10 4 помещали в каплях объёмом 20 мкл на крышку полистироловой чашки Петри с покрытием из силиконового эластомера СИЭЛ 159-330 (АО «ГНИИ-ХТЭОС», г. Москва, Россия) или без покрытия. В исследовании тестировали по три концентрации МЦ (0,1 %, 0,25 % и 0,4 %) и коллагена (150 мкг/мл, 300 мкг/мл и 600 мкг/мл). Скорость формирования клеточных конгломератов оценивали через изменение их площади спустя 4, 24, 48 и 72 часа культивирования. Результаты. Применение покрытия из СИЭЛ 159-330 позволило получить сфероиды таких же размеров, что и добавление 0,4 % МЦ на временном промежутке 72 часа. Силиконовое покрытие дополнительно уменьшило размеры клеточных сфероидов в среде с 0,1 % МЦ во всех временных точках, однако с ростом концентрации МЦ данный эффект исчезал. Кроме того, использование СИЭЛ 159-330 уменьшило связь размеров клеточных сфероидов с концентрацией МЦ, что позволяет рассматривать применение данного покрытия, как альтернативу МЦ или способ сократить её концентрацию. В опыте с добавлением в среду культивирования коллагена размеры клеточных конгломератов, образующихся на силиконовом покрытии, были достоверно меньше, чем на пластике без покрытия во всех вариантах опыта и временных точках. При этом эффект был более выраженным для концентрации коллагена 600 мкг/мл. Применение покрытия из СИЭЛ 159-330, кроме того, сократило вариативность размеров и формы образующихся клеточных конгломератов. Заключение. Ускоренная агрегация клеток и волокон внеклеточного матрикса в висячих каплях, а также сокращение вариативности в размерах и форме образующихся клеточных скоплений на СИЭЛ 159-330 позволяет сократить время проведения экспериментов и материальные затраты, как в опытах с добавлением веществ, ускоряющих формирование сфероидов (МЦ и коллаген), так и в их отсутствие.
Secondary plant metabolites are a promising source of anticancer drugs. Of particular interest are substances that can reduce the migration activity of cancer cells as potential anti-metastatic drugs. In present work we study the influence of 2,4-dihydroxy-2,5-dimethylfuran-3(2H)-one, 5-(hydroxymethyl)furan-2-carbaldehyde and corinan obtained from plants of the genus Petastes sp. on scratch healing rate of permanent cancer lines PC3, A431, CaCo2, HeLa, and T98G. Cells were grown in DMEM medium supplemented with 10% FBS. To perform a scratch test, cells were planted in an amount of 1,5105 per well of a 24-well plate. After cell adhesion, a vertical scratch was applied to the cell monolayer with a plastic tip, after which the medium containing the test substances at a concentration of 40 M was added. Then, within 48 hours, photographing and determining the scratch area were carried out. The degree of scratch overgrowth was determined as the ratio of the difference between the scratch areas after 48 hours of cultivation and the scratch area at the initial moment, expressed as a percentage. As a result of the experiment, it was shown that 2,4-dihydroxy-2,5-dimethylfuran-3(2H)-one downregulates scratch healing rate in cultures A431, HeLa, T98G. The high sensitivity of A431 to corinan was also shown.
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