Using multiple parallel sequencing on Illumina platform, we identified eight microRNAs that showed significant opposite changes of gene expression in cells of the hormone-sensitive LNCaP prostate cancer cell line and in cells of the hormone-resistant DU-145 cell line, in comparison to the microRNA expression in the normal prostate tissue cells. We found that the insulin-like growth factor 1 receptor (IGF1R) gene is a target of five microRNAs whose expression is increased in LNCaP cells and reduced in DU-145 cells.

We studied microRNA gene expression in HeLa cells following exposure for 6 h and 8 days to Co60 gamma rays at a dose of 4 Gy using an approach of large-scale parallel DNA sequencing. We identified 12 microRNAs with aberrant expression which were maintained in cell generations. The analysis of radiation-induced aberrant expression of pre-microRNAs made it possible to assess the importance of nuclear and cytoplasmic stages of microRNA biogenesis for preservation of its aberrant expression. On cell treatment by 5-azacytidine, aberrant expression was maintained only in two microRNAs: miR-21-3p and miR-422a, which demonstrated an increase in expression. Radiation-induced decrease in expression in ten examined microRNAs was dependent on DNA demethylation. At the same time, expression in a microRNA set, which demonstrated inheritable alteration of the expression after gamma-radiation exposure in the untreated cells, was not dependent or was weakly dependent on DNA methylation. The obtained results suggest that ionizing radiation induces aberrant DNA methylation, which affects inherited expression changes in microRNAs in cell generations after exposure to the mutagen.

It was first shown that DNA damage induction in mitomycin C-treated HeLa cells leads to a change in the selection of 5p and 3p microRNA duplex strands in the formation of the RNA-induced silencing complex (RISC).

Впервые показано, что индукция повреждений ДНК при обработке клеток HeLa митомицином C приводит к изменению селекции нитей 5р и 3р дуплекса микроРНК при формировании комплекса RISC (RNA-induced silencing complex).

The dependence of expression of miRNAs and their precursors (pre-miRNAs) on the DNA methylation level in HeLa cells 8 days after mitomycin C treatment was studied. A massive parallel DNA sequencing method was applied to analyze miRNA expression. 5-Azacytidine (DNA methylation inhibitor) was added to the medium 6 days after mutagenic agent exposure. The results indicated that the change in expression for some mature miRNAs (39 of 61) was accompanied by the change in the expression of their pre-miRNAs, while there were no significant changes in the expression of pre-miRNA for other mature miRNAs (22 of 61). The aberrant expression was maintained by 8 of 61 mature miRNAs and 6 of 55 pre-miRNAs in the induced HeLa cells after 5-azacytidine treatment. In addition, the expression of more than 90% of miRNAs, which indicated a significant change in expression after mitomycin C treatment, does not depend or depends slightly on the DNA methylation level in HeLa cells without mitomycin C treatment. The results suggest that mitomycin C induces aberrant DNA methylation which affects maintenance of changes in the miRNA expression in cell generations after mutagen treatment.

Purpose: to study in patients with chronic hepatitis C the pathogenesis hematopoietic activities of autologous blood hemolysate.Materials and methods: we made an indirect MALD-TOF-mass spectrometry of blood plasma in 37 patients with chronic hepatitis C who received the autologous blood hemolysate. The study was performed twice: before autohemotherapy and after 10 days of daily intravenous injections of 20.0 ml of the autologous blood hemolysate.Results: succeeded in isolating a blood plasma over 300 polypeptide sequence. Thus, as a result of intravenous administration of 20.0 ml autologous blood hemolysate frequency of IL-1β, IL-3, IL-6, IL-9, IL-11 and erythroferron significantly increased and the IL-10 and IL-13, by contrast, dropped .Summary: identification of proteomic changes in the profile of blood plasma in the predominance of proteins that stimulate hematopoiesis probably accounts for the clinical effectiveness of the autologous blood hemolysate.

Abstract.The aim of present study was complex evaluation and analysis of hormone and immune profiles in the patients with subclinical and manifesting thyroid hypofunction, taking into account a therapeutic drug monitoring for optimization of levothyroxine treatment. Forty patients with thyroid hypofunction are included into the study: I group, patients with subclinical hypothyreosis (n=19) and the levels of thyroxine-binding globuline exceeding 700 nmol/l, who received levothyroxine at 50 μg daily after withdrawal of drugs for 10 days; II group, the patients with manifesting hypothyreosis (n=21), who received levothyroxine at 100 μg daily. A significant regress of clinical signs of hypothyreosis was registered in both groups following 3 months of receiving levothyroxine, employing an optimized mode of dosage. An immune/endocrine component is shown for the mechanism of levothyroxine action, which promotes the development of maximal hormonal effect, with respect to improvement of life quality in the patients with hypothyreosis upon optimization of therapy based on the therapeutic drug monitoring using liquid chromatography/mass-spectrometry.

We identified 40 miRNAs with inherited aberrant expression by multiple parallel sequencing of human HeLa cells irradiated with X rays and mitomycin C. Twenty-two miRNAs were repressed and 15 miRNAs were induced after radiation and mytomycin C treatment. The expression of three miRNAs (miR-10b-5p, miR-148a-3p, and miR-340-5p) decreased after X-ray exposure and increased after mitomycin C treatment. The spectrum of aberrantly expressed miRNAs after X-ray and mitomycin C treatment is different, except for three miRNAs (mir-100-5p, miR-99b-5p, miR-501-3p), which showed the inherited decreased expression after both mutagens. It has been ascertained that for five miRNAs (miR-21-3p, miR-182-5p, miR-19b-3p, miR-30a-3p, and miR-30e-3p) with increased inherited expression, the targets are well-described tumor suppressor genes. For 9 miRNAs (miR-99b-5p, miR-148a-3p, miR-365a-3p, miR-193a-3p, miR-100-5p, miR-99a-5p, miR-29b-3p, miR-340-5p, and miR-23b-3p) with reduced inherited expression, the targets are oncogenes. The obtained results provide further support of the idea that induced epigenetic changes in the genome should be considered when assessing the long-term genetic effects of ionizing radiation and chemical compounds.

Applying the method of multiple parallel sequencing on the MiSeq platform (Illumina, United States), a comparative analysis of miRNA expression in tumor and normal colon tissue cells was performed. Forty miRNAs aberrantly expressed in cancer were detected. Among them, 15 and 25 miRNAs showed increased and decreased expression, respectively, for all or most of the cases. Sixteen miRNA clusters were identified, which showed a coordinated or incompletely coordinated aberrant expression in colorectal cancer cells. In two (miR-183/182 and miR-106b/25) and four (miR-143/145, miR-497/195, miR-30e/30c-1, and miR-30a/30c-2) miRNA clusters, respectively, a statistically significant coordinated increase or decrease in expression was registered for all miRNAs within the corresponding cluster. Three aberrantly expressed well-known miRNAs (miR-100-5p, miR-30d-5p, and miR-204-5p) were identified, which, however, had never before been associated with colorectal cancer. The obtained results demonstrate the potential and promising application of 6 miRNA clusters with coordinated aberrant expression as markers for colorectal cancer.

С помощью параллельного множественного секвенирования нуклеотидной последовательности проведен сравнительный анализ экспрессии микроРНК в клетках опухолевой и нормальной ткани толстой кишки и выявлено 40 микроРНК, аберрантно экспрессирующихся при развитии рака. Обнаружено увеличение экспрессии для 15 микроРНК и уменьшение экспрессии для 25 микроРНК. Идентифицировано 16 кластеров микроРНК с координированной или неполностью координированной аберрантной экспрессией в клетках опухоли толстой кишки. В двух (miR-183/182, miR-106b/25) и четырех (miR-143/145, miR-497/195, miR-30e/30c-1, miR-30a/30c-2) кластерах наблюдалось статистически значимое координированное увеличение и уменьшение экспрессии соответственно для всех микроРНК, входящих в кластер. Выявлено три аберрантно экспрессирующихся микроРНК (miR-100-5p, miR-30d-5p, miR-204-5p), для которых, однако, ранее не была показана связь с развитием рака толстой кишки. Полученные в работе результаты указывают на возможность использования шести координированно экспрессирующихся кластеров микроРНК в качестве маркеров развития рака толстой кишки.

С помощью метода множественного параллельного секвенирования идентифицированы микроРНК с наследуемыми изменениями уровня экспрессии в клетках HeLa после действия -квантов или митомицина С. Обнаружено снижение экспрессии 22 микроРНК и повышение экспрессии 15 микроРНК. Для трех микроРНК (miR-10b-5p, miR-148a-3p, miR-340-5p) выявлено повышение экспрессии при действии -квантов и уменьшение после действия митомицина С. Спектр аберрантно экспрессирующихся микроРНК при действии -квантов и митомицина С различается, за исключением трех микроРНК (mir-100-5p, miR-99b-5p, miR-501-3p), наследуемая экспрессия которых уменьшается как после -облучения, так и после обработки клеток митоцином С. В случае пяти микроРНК (miR-21-3p, miR-182-5p, miR-19b-3p, miR-30a-3p и miR-30e-3p) с увеличенной наследуемой экспрессией мишенями являются гены-супрессоры опухоли. Для девяти микроРНК (miR-99b-5p, miR-148a-3p, miR-365a-3p, miR-193a-3p, miR-100-5p, miR-99a-5p, miR-29b-3p, miR-340-5p и miR-23b-3p) с пониженной наследуемой экспрессией мишенями являются онкогены. Полученные результаты являются еще одним свидетельством того, что при оценке отдаленных генетических последствий эффектов ионизирующей радиации и химических соединений необходимо учитывать индуцированные эпигенетические изменения, связанные с функционированием микроРНК.

Using affinity chromatography, two-dimensional electrophoresis, and MALDI-TOF mass spectrometry, plasminogen isoforms were separated and identified in blood plasma. Healthy donors and patients with prostate cancer in various stages of development were included in the studied sample. With the development of prostate cancer, four additional specific plasminogen isoforms are registered in blood plasma; they are characterized by lower molecular weights and higher pI values compared to isoforms found in the control group.

According to some studies, the secretion of urinary glycoprotein Tamm‐Horsfall protein (uromodulin) plays a significant part in the suppression of calcium nephrolith formation. The aim of the present study was to detect correlation of Tamm‐Horsfall protein with soluble Ca2+ in the urine of urolithic patients and compare it with a control group, in order to estimate the degree of the disease progress, as well as the prognosis for the disease. The research included 29 urolithic patients, aged 8, 0±0, 2 y. The content of soluble Ca2+ in fresh urine was estimated by the method of capillary zone electrophoresis. The level of Tamm‐Horsfall protein in urine was measured from cryoprecipitate using SDS polyacrylamide electrophoresis and semiquantitative analysis. Urine from the urolithic patients was consistently higher in Ca2+ and in Tamm‐Horsfall protein concentrations than urine from controls, at all periods of the day.