E. Yu. Rykova

Serum immunoglobulins interact with oligonucleotides

The interaction of reactive derivatives of oligonucleotides bearing a 4‐[(N‐2‐chloroethyl‐N‐methyl)amino]benzylamin residue at the 5′‐terminal phosphate with serum blood proteins has been investigated. It was found that the compounds react with serum albumin and immunoglobulins M and G, the reactivity increasing in the order: albumin < IgG < IgM. The reactions with immunoglobulins were inhibited in the presence of different oligonucleotides, DNA and heparin, suggesting the oligonucleotide binding to some cationic region of the proteins. Myoglobin inhibited the interaction of oligonucleotide derivatives with myoglobin‐specific monoclonal antibodies which indicates that the derivatives interact with the proteins within or near the antigen binding site.

Plasma Content of Extracellular Nucleic Acids in Donors and Patients with Mammary Tumors

The concentrations of extracellular DNA and RNA were measured in the plasma of donors and patients with fibroadenoma and breast cancer. The content of extracellular DNA surpassed the normal in 80% plasma samples from patients with mammary tumors. Extracellular RNA was detected in 30% plasma samples from donors and patients with breast tumors. No correlations were found between plasma concentration of extracellular DNA and size and stage of tumor growth. Hence, measurement of extracellular DNA in the plasma of patients can be used only as an accessory test for tumor diagnosis.

Extracellular DNA in Culture of Primary and Transformed Cells, Infected and Not Infected with Mycoplasma

The composition and kinetics of accumulation of extracellular DNA in cultures of primary human endotheliocytes, cervical adenocarcinoma, and mycoplasma-infected cervical adenocarcinoma cells were studied. The content of DNA bound to cell surface did not change during culturing. The concentration of extracellular DNA in culture medium increased during the lag phase and at the beginning of the exponential growth phase, which probably attests to active secretion of DNA by cells. Spontaneous extracellular DNA synthesis was observed only in cell culture infected with mycoplasma.

Circulating microRNAs in lung cancer: Prospects for diagnosis, prognosis, and prediction of antitumor treatment efficacy

The review considers the main techniques to extract microRNA (miRNA) from various biological fluids (in particular, the serum and plasma), approaches to the analysis of miRNA concentration and composition, and methods to normalize the results in data analyses. Advantages and drawbacks of the methods are described. Special attention is given to circulating miRNAs, which can be used as markers for minimally invasive diagnosis, prediction of antitumor treatment efficacy, and disease prognosis in lung cancer. The review discusses the prospects and limitations that arise as the clinical significance is evaluated for miRNAs as potential tumor markers and a better understanding is gained for the roles various miRNAs play in the pathogenesis of lung cancer.

MIRA analysis of RARβ2 gene methylation in DNA circulating in the blood in lung cancer.

Analysis of DNA epigenetic mutations in the blood circulating DNA is a prospective trend for creation of noninvasive methods for the diagnosis and treatment effi ciency monitoring in cancer. The methylation status of target genes in circulating DNA was evaluated by methods based on preliminary bisulfi te conversion of DNA. We used a different approach based on selection of hypermethylated sequences of circulating DNA by means of DNA-methyl-binding protein (methylated CpG island recovery assay, MIRA). Methylation was evaluated for RARβ2 tumor suppression gene in circulating DNA in lung cancer and a trend was detected to higher methylation of this gene in the patients in comparison with healthy donors.

Interaction of keratin K1 with nucleic acids on the cell surface.

The interaction of surface proteins from A431 cells and cellular extracts with nucleic acids was investigated using affinity modification with 32P-labeled reactive oligonucleotide derivatives. Proteins with molecular weights of 68, 46, 38, and 28 kD as well as several low molecular weight proteins capable of binding to nucleic acids were found on the surface of intact cells. It was demonstrated that a protein with molecular weight of 68 kD is exposed at the cell surface, since the treatment of cells with trypsin results in the cleavage of this protein. Disruption of the integrity of the cell membrane (scrapping, treatment with trypsin, or permeabilization of the cell membrane with streptolysin O or saponin) disrupts the interaction of the reactive oligonucleotides with the cell surface proteins. Affinity modification of the cytosolic and membrane–cytosolic cell fractions with labeled oligonucleotides results in the modification of a large number of proteins, where proteins with molecular weights of 68, 46, 38, and 28 kD can be found as minor components. Surface oligonucleotide-binding proteins with molecular weight of ∼68 kD were isolated by affinity chromatography after the modification of intact A431 cells with a reactive oligonucleotide derivative. The isolated surface oligonucleotide-binding proteins from A431 cells were sequenced, and one of the proteins was identified as keratin K1.

Plasmid DNA Modulates Chronic Graft-versus-Host Reaction

Injection of the parental lymphoid cells to the F1 hybrids is known to result in either Th1-dependent acute or Th2-dependent chronic graft-versus-host reaction (GVHR). The clinical picture of the disease depends on genetic differences between the donor and the recipient, the number of transferred cells, the donor cell phenotype, and other factors. Inoculation of the hybrids ( C 57 BL /6 × DBA /2) F 1 ( B 6 D 2 F 1) with the DBA/2 cells led to a Th2-dependent chronic GVHR characterized by activation of donor Th-cells, polyclonal activation of recipient B lymphocytes, the production of autoantibodies, and development of lupuslike glomerulonephritis [1].

Molecular genetic markers in diagnosis of lung cancer

The review considers the main approaches to the identification of lung cancer (LC) markers, including genetic, epigenetic, protein, transcriptomic, proteomic, metabolic, and miRNA markers. Emphasis is placed on epigenetic markers, which are the most promising because epigenetic changes are among the earliest events in malignant transformation. Special attention is given to circulating tumor markers, which can be detected in easily accessible biological fluids with minimally invasive methods and may be useful for screening the risk groups for LC, diagnosing cancer before its clinical manifestation, monitoring the tumor in remission after therapy, and verifying the diagnosis based on standard clinical and instrumental methods of diagnostics. Extracellular nucleic acids (circulating in blood, circNA) are highlighted as a potential source of material for early diagnosis of LC, prediction of the efficiency of antitumor treatment, posttreatment monitoring, and disease prognosis.

Representation Analysis of miRNA in Urine Microvesicles and Cell-Free Urine in Prostate Diseases

Urine of prostate cancer patients contains tumor-specific biopolymers, including protein- and microvesicles-associated miRNAs that can potentially be used as oncomarkers. Previously we have characterized urinary extracellular vesicles and demonstrated diagnostic potential of their miRNAs. In this study, we have performed a comparative analysis of 84 miRNA in paired samples of urine microvesicles and urine supernatant from healthy men, patients with benign prostate hyperplasia, and prostate cancer patients using miRCURY LNA miRNA qPCR Panels. In all groups of patients, miRNA subsets characterized by different distribution between the urinary fractions have been found. In this context, two groups of miRNAs have been identified, which are involved in several signaling pathways including those associated with prostate cancer development.

Bioinformatics analysis for evaluation of the diagnostic potentialities of miR-19b, -125b and -205 as liquid biopsy markers of prostate cancer

Presence of tumor-derived cell-free miRNA in biological fluids as well as simplicity and robustness of cell-free miRNA quantification makes them suitable markers for cancer diagnostics. Based on previously published data demonstrating diagnostic potentialities of miR-205 in blood and miR-19b as well as miR-125b in urine of prostate cancer patients, bioinformatics analysis was carried out to follow their involvement in prostate cancer development and select additional miRNA-markers for prostate cancer diagnostics. Studied miRNAs are involved in different signaling pathways and regulate a number of genes involved in cancer development. Five of their targets (CCND1, BRAF, CCNE1, CCNE2, RAF1), according to the STRING database, act as part of the same signaling pathway. RAF1 is regulated by miR-19b and miR-125b, and it was shown to be involved in prostate cancer development by DIANA and STRING databases. Thus, other microRNAs regulating RAF1 expression such as miR-16, -195, -497, and -7 (suggested by DIANA, Ta...