The review of the literature about the results of the study of the role of 4a/b VNTR-polymorphism of eNOS in pathophysiology of various states of the body. It is shown that the data are ambiguous and sometimes contradictory. The study of the contribution of candidate genes to the implementation of multifactorial diseases can increase the accuracy of prediction of the set of risk factors, early diagnosis facilitate and sufficient therapy of multifactorial diseases.

Scorpion venom is a complex multicomponent mixture of biologically active substances, some of which possess very interesting properties and are used in quite unexpected fields. The family of chlorotoxin (CTX)-like peptides serves a good example. These toxins exhibit insecticidal activity, however, their molecular mechanism of action on insect organism remains elusive. Nevertheless, CTX-like peptides attracted considerable research effort due to their ability to specifically interact with cells of brain tumors, i.e. gliomas. In the future these compounds may considerably aid anticancer therapy. This review summarizes the results obtained during the past 40 years of CTX-like peptides investigation. Both biological function aspects and the applied field related to gliomas are considered.

The participation of the main caspases in the cytotoxic effects induced by monoclonal antibody 14G2a specific against tumor-associated ganglioside GD2 was studied in the EL-4 cells. It has been found constitutive expression ofprocaspases genes in the EL-4 cells; incubation of the cells with 14G2a antibodies didnot result in increasing of the procaspases expression. Weak enzymatic activity of caspases has been shown using fluorescent labeled substrates. At the same cell death level, activity of caspase-3 and caspase-9 in the cells incubated with 14G2a was about 7.5- and 3-fold lower than in cells after incubation with staurosporine. Pan caspase inhibitor Z-VAD-FMK, and caspase-3 inhibitor reduced the cytotoxic effects induced by 14G2a at 9-16 and 6-13%, respectively. At the same conditions, pan caspase inhibitor decreased staurosporine-induced apoptosis at 55-65%. Inhibitors of other caspases had no effect on the cell death triggered by the antibodies. Inhibition analysis demonstrated also that caspases did not involved in the cell volume decreasing and permeabilization of the cell plasma membrane, which were the first stages of anti-GD2-mAb-induced cell death in the EL-4 cells. Thus, despite the slight activation of caspases during the cell death induced by antibodies directed to GD2, they do not play a key role and do not determine the mechanism of cell death triggered through the tumor-associated ganglioside GD2.

A key role in tumor progression play two processes--the destruction and angiogenesis. Matrix metalloproteases (MMPs) play a leading role during tissue degradation. Tissue collagenase--MMP-1 and MT1-MMP hydrolyze fibrillar collagens, which are the basis of connective tissue matrix, and ensure the development of an invasive process. Gelatinase A and B (MMP-2 and MMP-9) hydrolyze collagen type IV, which is the basis of the basal membrane, and facilitate the development of metastasis. Endogenous tissue inhibitors TIMP-1 and TIMP-2 are involved in the regulation of MMP expression and activity. It has been established that MMP-9 release vascular endothelial growth factor (VEGF) associated with the STM--the primary inductor angiogenesis. Angiotensin-converting enzyme (ACE) participates in the induction of VEGF synthesis. ACE--a key enzyme of the renin-angiotensin system, forms angiotensin II, which interactes with the receptor ATIR and induces VEGF synthesis, as well as stimulates endothelial cell proliferation. Our experimental studies devoted to the study of particularity expression of key enzymes of destruction and angiogenesis in squamous cell carcinoma of the cervix (SCC). It was studied: MMP-1, MT1-MMP, MMP-2 and MMP-9 and their endogenous regulators: TIMP-1, TIMP-2, and as well as ACE. Work was performed on clinical specimens containing the tumor tissue, taking into account the presence or absence of metastasis to regional lymph nodes and the specimens of adjacent morphologically normal tissue. It was shown that the increase of MMP-1, MT1-MMP and MMP-9 expression and low of TIMP-1 and TIMP-2 expression makes the main contribution to the destructive (invasive) potential of SCC. The change of MMP-2 expression is not so significant and it is less influenced to the destructive potential. It was shown dramatic increasing of MMP-1 and MMP-9 activity in metastasizing tumor tissue ACE activity in a tumor in most of the samples was higher than the activity in normal tissues. It was established that the expression of key enzymes degradation and angiogenesis occurs not only in tumor but also in normal tissues. Data are important for understanding the mechanisms of tumor progression and have prognostic value and may affect the therapeutic strategy for patients.

The ability to active movement in extracellular matrix wherein significant role plays remodeling of the cytoskeleton by actin-binding proteins may influence on the metastatic potential of tumor cells. We studied the expression of actin-binding proteins and β-catenin in connection with proteasome and calpain functioning in the tissues of primary tumors and metastases of ovarian cancer. The chymotrypsin-like proteasome activity and calpain activity were shown to be significantly higher in ovarian cancer than in normal tissues. Furthermore, the activity of the proteasome and calpain were significantly higher in the peritoneal metastases in comparison with primary tumors. Correlation analysis showed in the primary tumor tissue the presence of a positive relationship between the activity of calpain and chymotrypsin-like proteasome activity (r = 0.82; p = 0.0005), whereas in metastases this connection was not revealed. Contents of p45 Ser β-catenin and the actin-severing protein gelzolin were decreased in metastases relative to primary tumors. Level of cofilin, functionally similar to gelzolin protein, was significantly higher in metastases compared to primary ovarian tumor tissue. In ovarian cancer significant reduction in the number of the monomer binder protein thymosin-β4 was observed in primary tumors and metastases as compared to normal tissues, but significant differences between the primary tumor and metastases were not observed. In the tissues of primary tumors negative correlations were observed between the chymotrypsin-like activity of the proteasome and the amount of p45 Ser β-catenin and protein Arp3, a member of the Arp2/3 complex. In metastasis negative correlation were revealed between the activity of calpain and content Arp3, cofilin, thymosin. The data obtained suggest the existence of different mechanisms of proteolytic regulation of locomotor proteins in primary tumors and metastases in ovarian cancer.

Proteasomes carry out regulated proteolysis of most proteins and thereby play a crucial role in the regulation of different cellular processes. Dissecting subunit composition and post-translational modifications of proteasome is one of the important milestones in understanding their functions and mechanisms of regulation in the cell. To this end a strategy we followed a strategy for affinity purification of proteasomes from human myeloid leukemia cells with subsequent mass spectrometric analysis. Proteasomes were purified from the stable cell line K562 expressing HTBH tag-labeled 20S proteasome subunit β7 (PSMB4) by non-covalent affinity purification on biotin-avidin beads, followed by elution with TEV protease. We identified all known subunits of the 26S proteasome, as well as PA200 and regulators PA28γ amongst the eluted proteins, using MALDI-ICR mass spectrometry. We have shown that the proteasomes are associated with heat shock proteins, components of the ubiquitin-proteasome system of some cytoskeleton proteins. A number of novel phosphorylation, ubiquitination and N-terminal modification of proteasome subunits were found for 16 proteasome subunits. Our results might be useful for further proteomic studies of proteasomes.

The expression of the total proteasome pool, immune proteasome subunits LMP2 and LMP7, TAP1 and TAP2 transporters, as well as RT1A molecule of MHC class I was investigated in the ascite Zajdela hepatoma at the 10th day after implantation into Brattleboro rats with the hereditary defect of hypothalamic arginine-vasopressin synthesis (AVP) and into WAG rats with normal AVP expression. In Zajdela hepatoma cells implanted into Brattleboro rats the 3-fold increase of the total proteasome pool and LMP2 level and 8-fold increase of the LMP7 level was detected by Western blotting as compared to those in WAG rats. Differences in the LMP2 and LMP7 expression suggest variations in their functions, namely the important role of LMP7 in anti-tumor immunity. The growth of Zajdela hepatoma in WAG rats was accompanied by the decreased level of total proteasome pool as well as immune proteasome expression as compared to those in Brattleboro rats during the regression of tumor. The analysis of TAP1 and TAP2 revealed the pronounced expression of these peptide transporters in Zajdela hepatoma cells implanted into Brattleboro and WAG rats. The expression level of RT1A molecule of MHC class I was increased 3 times in Zajdela hepatoma cells implanted into Brattleboro rats as compared to WAG rats. Moreover, flow cytometric analysis of CD4- and CD8-lymphocytes number in the spleen of Brattleboro and WAG rats was performed at the 10th day after implantation of Zajdela hepatoma. The increased number of CD4- and CD8-lymphocytes was observed in the spleen of Brattleboro as compared to WAG. The increased subpopulations of cytotoxic T-lymphocytes and T-helpers might promote the tumor regression in Brattleboro rats. The reduced populations of CD4- and CD8-lymphocytes in the spleen of WAG rats were accompanied by the splenomegaly and tumor progression. The data obtained suggest that AVP deficiency in Brattleboro rats leads to the increase of the immune proteasome and MHC class I expression in Zajdela hepatoma cells, resulting in tumor immunogenicity and its elimination by the adaptive immunity.

The main physiological function of plasmin is a blood clot fibrinolysis and restore normal blood flow. To date, however, it became apparent that in addition to thrombolysis plasminogen/plasmin system plays an important physiological and pathological role in the degradation of extracellular matrix, embryogenesis, cell migration, tissue remodeling, wound healing, angiogenesis, inflammation and tumor cells migration. This review focuses on the structural features of plasminogen, the regulation of its activation by physiological plasminogen activators, inhibitors of plasmin and plasminogen activators, the role of the plasminogen binding to fibrin, cellular receptors and extracellular ligands in performing various functions by formed plasmin.

The paper gives general information about the epithelial-mesenchymal transition (EMT), its morphological manifestations, altered expression of a number of proteins, types of EMT, and its role in embryogenesis and human diseases, including that about EMT as a mechanism by which the tumor cell acquires prometastatic potential. EMT is a process that is essential in health for gastrulation and the formation of neural crest cells; however, it is also important for the development of abnormalities, among other processes, organ fibrosis and tumor metastases. An understanding of the role of EMT in cancer spread has led to active studies of the process in the past decades. Despite the fact that there are sufficiently many publications on different aspects of EMT, the exact mechanisms regulating the process and the possibility for its therapeutic exposure remain unclear. There is also evidence on the possible association of EMT with the generation of cancer stem cells in tumors.

According to current views, high-grade prostatic intraepithelial neoplasia is the most likely precursor of prostate adenocarcinoma. This review gives the latest data of genetic, proteomic, and morphological analyses of this neoplasia and touches upon the probems that might arise when searching for new markers for differential diagnosis and prognosis estimation.

The paper characterizes adrenocortical oncocytoma, a rare adrenal tumor, accompanied by Cushing's syndrome and estrogen and androgen production and provides histological and immunohistochemical features. The authors describe their observation of a 33-year-old female woman. It is shown that estimation of the malignant potential of adrenocortical oncocytomas requires a special approach and must be done using the Lin-Weiss-Bisceglia criteria.

To estimate changes in the trend of growth of primary tumor nodules, the degree of lymphocytic infiltration, and the expression levels of oncomicroRNA miR-21 and miR-let-7b when inhibiting matrix metalloproteinases 9 and 13 (MMP-9 and MMP-13) in vivo in C57B16 mice with transplantable melanoma B-16.

To study a relationship of survival rates to the expression of major genes (p53, PTEN, COX2, pRb and others) involved in the pathogenesis of ovarian cancer.

Germline mutations of BRCA1/2 genes cause the predisposition of their carriers to breast or/and ovary cancers (BC or/and OC) during the lifetime. Identification of these mutations is a basis of molecular diagnosis for BC susceptibility. Rapid genotyping technique using microarrays for identification of BRCA1 185delAG, 300T>G, 4153delA, 5382insC mutations and 4158 A>G sequence variant; BRCA2 695insT and 6174delT mutations; 1100delC mutation in CHEK2 gene was applied for 412 randomly collected breast cancer samples from the central region of European area of Russia. In 25 (6.0%) patients (6.0%) BC was associated with other tumours: OC, cervical cancer, colorectal cancer etc. BRCA1/2 and CHEK2 mutations were found in 33 (8.0%) BC patients. The most frequent mutation was BRCA1 5382insC, occurred in 16 (3.9%) BC patients, and CHEK2 1100delC, revealed in 7 (1.7%) BC patients. An application of diagnostic BC-microarray for genetic testing of BRCA1/2 and CHEK2 founder mutations has been discussed.

Review is devoted to analysis of the role of microRNA in progression of human tumors. The following aspects of this problem are discussed: general characteristics of microRNA, expression pattern of these RNAs in human tumors and specificity of this expression, putative role of microRNAs as oncogenes and tumor suppressors for tumor growth, participation of microRNAs in induction of transformed phenotypes of tumor cells, possible role of microRNAs for early diagnosis of the disease and prognosis.

Some ribonucleases (RNases) produce selective toxic effect on the cancer cells. The mechanism of this antitumor activity remains largely unclear. The subject of this review is the RNases interaction with cellular components, resulting in the induction of apoptosis of tumor cells. Cell surface structures, which are potential acceptors of the exogenous RNase are discussed: acidic lipids and glycoproteins, heparansulfate-containing proteoglycans, actin, and RNA-associated proteins. Cell membranes of normal and malignant cells differ according to the composition of these components, which largely determines the selectivity of RNases for the latter. Different types of RNA are examined as intracellular targets of the RNases activity, evidence is presented demonstrating the possibility of exogenous RNases intervening in the process of RNA interference. The role of potassium channels, NF-kappaB-dependent.signaling pathway and various caspases in apoptosis induced by exogenous RNases is discussed. Evidence is also presented showing that the sensitivity of cells to exogenous RNases is linked to the expression of certain oncogenes, namely RAS, KIT, AML1-ETO. It is suggested that discovering the details of the mechanisms of RNases cytotoxic effect in malignant cells susceptible to their activity, will in the future serve as a foundation to developing new tools of targeted anticancer therapy.

Recent data obtained at the junction of biochem: istry, molecular and cell biology and experimental oncology, showed that the formation of secondary foci of tumor growth during cancer progression--metastasis formation--is a highly determinate and regulated process. This process includes on the one hand the appearance of metastatic population of cells with special characteristics that allow their dissemination and seeding in distant organs and on the other.hand the formation of specific attractive micro environment in target organs. These cells show the ability to switch their motility to the most effective mode depending on the properties of the surrounding tissues (plasticity), appearance of specific receptors on the cell surface, which enhance their directed migration to target organs and acquisition of some characteristics of stem cells, allowing them to survive and reproduce in alien microenvironment. These alterations are strongly coordinated with development of a specific nichein the target organ which stimulates initiation and growth of a future metastasis, so-called premetastatic niche. In this review we analyzed recent data concerning mechanisms which regulate the emergence of metastatic population of cells, development of premetastatic niches and coordination of these processes:.

The genetic characteristics are key risk factors of development of many human neoplasms including B-cell tumors of lymphatic system. The relationship between polymorphic variants of genes FCGR2A (His 1 66Agr), CD14 (C-159T). IL1β (T-31C), IL2 (7:330G) and 7LR2 (Arg753Ghn) and development of various forms of B-cell tumors of lymphatic system in 80 patients was investigated. The statistically significant differences of rates of particiular genotypes of single nucleotid polymorphisms of genes FCGR2A, CD14. IL1β, IL2 and TLR2 in patients with indolent and aggressive types of course of non-Hodgkin lymphoma and also multiple myeloma. The results prove hypothesis that genetic variants of genes of inborn immune response effect the origin and character of course of different types of lymphoproliferative diseases. The markers can become additional prognostic characteristics of benign and aggressive course of tumors.

A recent study of human normal and tumor tissues revealed a high transcriptional activity of pericentromeric satellite DNA repeats (they produce half of all transcripts in tumor cells that is many times higher than in normal ones). It was found also that the two subtypes of satellite DNA (HSATII and GSATII) are transcribed reciprocally, i.e. there is a sharp prevalence of HSATII transcription in tumors, while GSATII transcription prevails in the corresponding normal tissues. As different RNAs are present in blood plasma, and some of them serve as effectivetumor markers, we attempted for the first time to evaluate satellite HSATII and GSATII RNAs in the blood plasma of healthy donors and cancer patients. The RT-PCR protocol designed for this purpose allowed us to detect transcripts of both HSATII and GSATII repeats. As it has been shown, HSATII transcripts are more abundant than GSATII ones in plasma of healthy donors and vice versa in plasma of cancer patients; these ratios being diametrically opposed to those that exist within the cells. Some suggestions concerning origins of circulating satellite RNAs and their probable role as tumor markers are discussed.

Since the discovery of potential therapeutic value of quadruplex secondary nucleic acids structures, many compounds that stabilize these targets were found. Such progress became possible due to understanding of the structural features of G-quadruplexes. Quadruplex ligands selectively suppress the growth of tumor cells by indirect inhibition of the telomerase activity and/or attenuation of oncogenes' overexpression. Therapeutic effect demonstrated in vivo supports the prospect of such compounds for the development of the targeted anticancer drugs. This review reveals the significance of G-quadruplexes as therapeutic targets and focuses on biochemical properties of the low molecular weight quadruplex ligands.