Ejc Supplements最新文献

{"title":"P124","authors":"A. Bondar , A. Kurilshikov , E. Morozkin , M. Zaripov , M. Kabilov , V. Voytsitskiy , V. Vlassov , P. Laktionov","doi":"10.1016/j.ejcsup.2015.08.012","DOIUrl":"10.1016/j.ejcsup.2015.08.012","url":null,"abstract":"<div><p>Cancer cells display altered methylation signatures distinguishing them from normal cells. Originating from all tissues and cells of the body cell-free DNA (cfDNA) including aberrantly methylated DNA reflect epigenetic aberrations occurs not only in tumor cells but also in tumor microenvironment. Actually, aberrantly methylated cfDNA has proved to be a promising biomarker for noninvasive detection of cancer with several clinically-certified tests (Epi-pro Colon®, Epi-pro Lung®, Cologuard®). However only one test (Epi-pro Colon®) uses blood plasma – the most convenient source of cfDNA. Input of tissue and age specific methylation along with unidentified reasons lead to the presence of molecules with every conceivable cytosine-methylation patterns which decrease probability of tumor DNA identification. Among numerous variants of cfDNA methylation patterns only few reflect cancer related changes. To identify those tumor-specific profiles single nucleotide resolution of methylated cytosine locations in the individual circulating DNA molecules is obviously required.</p><p>We performed target bisulfite sequencing of potential prostate cancer (PC) cfDNA markers (GSTP1; RNF219) isolated from blood plasma of 18 healthy donors (HD), 17 benign hyperplasia (BPH) and 20 PC patients using MiSeq platform (Illumina). RNF219 gene was shown to have high diagnostic potential in our previous comparative study of cfDNA from HD, PC and BHP patients using HCGI12k microarrays (Cortese, et al., 2012). GSTP1 is a common pathological DNA methylation event in PC and is most widely studied and promising methylation marker in the cfDNA of PC patients (Wu, et al., 2011).</p><p>Selected loci were amplified after bisulfite conversion (Zymo Research) of cfDNA with methyl-independent barcoded primers and sequenced with coverage ranging from 23509 to 143953. Identification of CpG methylation status in DNA fragments was performed with the BiQ Analyzer HT Software. All statistical analysis was performed using R Statistical Software (version 3.1.1) To reveal diagnostically significant differences, several approaches to data analysis were used. Conventional approach is a prediction of patient’s diagnosis based on differences in methylation level of CpG-sites. Another approach used in this study relies on discrimination of cancer-related correlation between methylation statuses of CpG-sites within individual molecules of cfDNA – Intramolecular Correlation of Methylation Statuses (ICoMS).</p><p>Study population was randomly subsampled into training and test cohorts. The logit regression model based on methylation level of CpG-sites achieved an area under the ROC curve (AUC) exceeding 0.94 in both cohorts for GSTP1 gene and 0.81 – for RNF219 gene. A novel approach to identify diagnostic significance of cfDNA methylation was based on comparison of correlation matrices (pairwise phi coefficient between methylation statuses of CpG sites) for HD, BHP and PC groups. Binomial regression","PeriodicalId":11675,"journal":{"name":"Ejc Supplements","volume":"13 1","pages":"Page 7"},"PeriodicalIF":0.0,"publicationDate":"2015-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.ejcsup.2015.08.012","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54308744","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"P132","authors":"M. Chernyavskaya , V. Chernych , A. Efremov , V. Atamanov","doi":"10.1016/j.ejcsup.2015.08.019","DOIUrl":"10.1016/j.ejcsup.2015.08.019","url":null,"abstract":"<div><h3>Purpose</h3><p>To research the characteristics of the inflammation immune disorders and cell destruction at the local level in the lacrimal and intraocular liquids in patients with choroidal melanoma.</p></div><div><h3>Materials and methods</h3><p>Investigation of concentration of the cytokines interleukin (IL) IL-4, IL-6, IL-8, IL-10, autoantibodies to antigens of native DNA (AAB to Ag of nDNA) in the lacrimal and intraocular liquids was performed in 36 patients (72 eyes) aged 34–83<!--> <!-->years. The mean age of the study group patients was 60.45<!--> <!-->years (15 males and 21 females). As standard indicators, survey results of 20 “healthy” donor lacrimal liquid – volunteers were used. For statistical data processing, descriptive statistics and methods of inter-group comparisons were used. Quantitative characteristics are presented as the median (25, 75 percentile). For comparison of quantitative traits, regression analysis was used. <em>P</em> value of <0.05 considered significant (95% confidence interval). Statistical analysis was performed using the software R (Team RC, 2013).</p></div><div><h3>Results</h3><p>A significant positive relation of concentration of IL-4, IL-6, IL-8, IL-10, AAB to Ag of nDNA in lacrimal liquid of eye with choroidal melanoma and paired “healthy” eye was reveled (<em>p</em> <!--><<!--> <!-->0.05). Concentrations of IL-6, IL-8, IL-10 and AAB to Ag of nDNA in the lacrimal liquid of the eye with choroidal melanoma were significantly higher as compared with the levels of the control group (<em>p</em> <!--><<!--> <!-->0.05). The level concentration of IL-4 in lacrimal liquid in eye with choroidal melanoma was significantly low (<em>p</em> <!--><<!--> <!-->0.05). The level concentration of IL-6 was significantly higher in the early (T1–T2) than in the later stages (T3–T4) of choroidal melanoma development (<em>p</em> <!--><<!--> <!-->0.05). All stages of choroidal melanoma were characterized by higher level of IL-6, compared with the control group (<em>p</em> <!--><<!--> <!-->0.05). Given the role of IL-6 in the pathogenesis of malignant diseases, the activity may be indicative of tumor progression in the early stages of development of choroidal melanoma. A significant increase in IL-10 was shown at stages T1 and T4 compared with the control group (<em>p</em> <!--><<!--> <!-->0.05). Regression analysis revealed a significant positive relation of concentration of IL-4, IL-6, IL-8, IL-10, AAB to Ag of nDNA between lacrimal liquid and intraocular liquids of the eye with choroidal melanoma (<em>p</em> <!--><<!--> <!-->0.05).IL-4 <em>r</em> <!-->=<!--> <!-->0,84 (0.58; 1.47), <em>p</em> <!--><<!--> <!-->0.05 IL-6 <em>r</em> <!-->=<!--> <!-->0.98 (0.64; 1.32), <em>p</em> <!--><<!--> <!-->0.01 IL-8 <em>r</em> <!-->=<!--> <!-->0.82 (0.12; 1.73), <em>p</em> <!-->=<!--> <!-->0.05 IL-10 <em>r</em> <!-->=<!--> <!-->0.84 (0.01; 1.81), <em>p</em> <!-->=<!--> <!-->0.05AAB to Ag of nDNA <em>r</e","PeriodicalId":11675,"journal":{"name":"Ejc Supplements","volume":"13 1","pages":"Page 11"},"PeriodicalIF":0.0,"publicationDate":"2015-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.ejcsup.2015.08.019","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54308865","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"T33","authors":"M. Freidin","doi":"10.1016/j.ejcsup.2015.08.029","DOIUrl":"10.1016/j.ejcsup.2015.08.029","url":null,"abstract":"<div><p>Blood-derived biomarkers, such as circulating tumour cells (CTCs) and circulating tumour DNA (ctDNA), are a valuable source of molecular genetic data for tumours they spring from. In translational cancer research, the “liquid biopsy” concept has been put forward to denote the detection and molecular characterization of these biomarkers.</p><p>The idea of liquid biopsy is based on a hypothesis that profiles of somatic mutations in CTCs and ctDNA are identical to those in the original tumour. Therefore, the mutation status of the source tumour can be revealed through the molecular analysis of the CTCs and ctDNA obtained from the blood. The major advantages of liquid biopsy are an essentially decreased invasiveness (no need for tissue biopsy, surgery or bronchoscopy) and an ability to carry out the analysis at patient’s follow up (e.g. to monitor for residual disease).</p><p>However, both the CTCs and ctDNA are not abundant in the bloodstream and their capture is technically challenging. Also, due to tumour heterogeneity, the CTCs may not fully represent the entire tumour, while ctDNA is naturally fragmented and degraded, so its utility for genetic analysis may be limited. Finally, the DNA extracted from CTCs and, especially, ctDNA are “contaminated” by DNA from non-tumour cells from the bloodstream raising a challenge of detecting mutant DNA among significantly prevailing wild-type DNA.</p><p>Some of these issues can be overcome by using such advanced techniques as BEAMing, digital PCR or ultra-deep sequencing, but their use in standard clinical settings is limited by the need for special equipment and associated costs. Inexpensive and less sophisticated, but still highly sensitive and specific, approaches, such as COLD-PCR or wild-type blocking PCR, are also available to detect “druggable” mutations in CTCs and ctDNA.</p><p>Application of these approaches of liquid biopsy in clinical practice may be highly beneficial for personalized care of lung cancer patients.</p></div>","PeriodicalId":11675,"journal":{"name":"Ejc Supplements","volume":"13 1","pages":"Page 16"},"PeriodicalIF":0.0,"publicationDate":"2015-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.ejcsup.2015.08.029","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54309046","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"P68","authors":"E. Grigorieva ,&nbsp;M. Karbyshev ,&nbsp;V. Volkomorov ,&nbsp;E. Kremmer ,&nbsp;A. Huber ,&nbsp;I. Mitrofanova ,&nbsp;M. Zavyalova ,&nbsp;E. Kaigorodova ,&nbsp;J. Kzhyshkowska ,&nbsp;N. Cherdyntseva","doi":"10.1016/j.ejcsup.2015.08.033","DOIUrl":"10.1016/j.ejcsup.2015.08.033","url":null,"abstract":"<div><h3>Background</h3><p>Transmembrane prostate androgen-induced protein 1 (TMEPAI) is a membrane protein that has attracted significant attention of many researchers its involvement in TGF-<em>β</em> signaling pathway which involved in malignant transformation and metastatic tumor progression. We investigated the TMEPAI expression level in gastric adenocarcinomas in comparison to non-tumor mucosa samples and determined its potential prognostic significance.</p></div><div><h3>Materials and methods</h3><p>Fresh and paraffin-embedded gastric adenocarcinoma samples and paired adjacent normal tissues were collected from gastric cancer patients. Evaluation of the PMEPA 1 gene expression was carried out using RT-PCR. For evaluation of TMEPAI protein expression, monoclonal antibodies (mAbs) were developed by using hybridoma techniques. Specificity of prepared monoclonal antibodies against recombinant TMEPAI and evaluation of its expression in the clinical samples using selected mAbs were performed using immunoblotting and immunohistochemistry.</p></div><div><h3>Results</h3><p>We have identified more than two-fold increase in gene expression of PMEPA1 in tumor tissue in 44% of patients. The monoclonal antibodies have shown the capacity to specifically recognize the recombinant TMEPAI in HEK293T cell lysates. We also evaluate the ability of the selected antibodies to recognize the target protein in fixed cells by immunocytochemistry. The evaluation of TMEPAI in adenocarcinoma samples collected from gastric cancer patients revealed decreased protein expression. We have observed pronounced expression of TMEPAI in normal gastric epithelial cells, while tumor cells from gastric adenomas and adenocarcinomas samples were mostly negative for target protein expression. We found that gastric epithelium cells lose the TMEPAI expression concurrent with severe dysplasia.</p></div><div><h3>Conclusion</h3><p>Apparently, the TMEPAI may be a potential biomarker of malignant transformation risk of the stomach epithelium.</p><p>The presented study was financially supported by Grants from the <span>Russian Fund for Basic Research</span> (<span>14-04-31500</span>) and Tomsk State University Competitiveness Improvement Program.</p></div>","PeriodicalId":11675,"journal":{"name":"Ejc Supplements","volume":"13 1","pages":"Pages 18-19"},"PeriodicalIF":0.0,"publicationDate":"2015-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.ejcsup.2015.08.033","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54309115","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"P64","authors":"T. Kunts ,&nbsp;K. Karpukhina ,&nbsp;E. Mikhailova ,&nbsp;N. Varaksin ,&nbsp;A. Autenshlyus","doi":"10.1016/j.ejcsup.2015.08.053","DOIUrl":"10.1016/j.ejcsup.2015.08.053","url":null,"abstract":"<div><p>Growing tumor and its microenvironment are capable to produce a number of cytokines that alter the nature of the antitumor surveillance by host immune system.</p></div><div><h3>Objective</h3><p>Comparative evaluation of cytokine-producing function of the invasive ductal carcinoma and fibroadenoma of the breast in vitro.</p></div><div><h3>Materials and methods</h3><p>Similar biopsies (<em>V</em> <!-->=<!--> <!-->8<!--> <!-->mm³) of the breast tumors were obtained using a special device, cultivated in DMEM F-12 at 37<!--> <!-->°C for 72<!--> <!-->h. Concentrations of the following cytokines: IL-1<em>β</em>, IL-1Ra, TNF<em>α</em>, IL-2, IL-6, IL-8, IL-10, IL-17, IL-18, VEGF and IFN<em>γ</em> in the supernatant of the tumor were measured with enzyme-linked immunosorbent assay (ELISA).</p></div><div><h3>Results</h3><p>The investigation of cytokines level in the supernatant of malignant and benign breast tumors revealed significant differences only in concentrations of IL-10, IL-17, IL-18 and IFN<em>γ</em> which had a contrary tendency. For example, the concentration of IL-10 was lower at invasive ductal carcinoma in comparison with fibroadenomas. The concentrations of IL-17, IL-18 and IFN<em>γ</em> at invasive ductal carcinoma were significantly higher than those of breast fibroadenomas. IL-17 and IL-18 are known to be pro-oncogenic cytokines, and the higher the level, the higher the severity of tumor progression. Reduction in the IL-10 concentration might be explained by the already formed neoplasm, which depends on angiogenesis. In this case, IL-10 no longer exerts antiangiogenic action, which contributes to tumor progression. Reduction in the IL-10 concentration, which inhibits the production of IFN<em>γ</em> leads to an increase in the IFN<em>γ</em> level. In early stages of tumor development, IFN<em>γ</em> provides an antitumor effect and at the same time facilitates the selection of a more malignant clones but its pro-tumoral action predominates at advanced stages of tumorigenesis. Moreover, higher concentration of IFN<em>γ</em> is supposed to be associated with biological effects of IL-18, which is its immediate inductor. In addition, malignant tumor cells are capable to produce their own IL-18, which stimulates tumor progression and facilitates the migration of endothelial cells involved in angiogenesis, which leads to intensified invasion and metastasis.</p></div><div><h3>Conclusion</h3><p>Cytokine production in supernatants of invasive ductal carcinoma compared with fibroadenoma of the breast is characterized by increase in IL-17, IL-18 and IFN<em>γ</em> concentrations and decrease in IL-10 concentration. Findings suggest the ability of malignant tumor and its microenvironment to secrete the pro-oncogenic cytokines. Fibroadenoma is also able to produce cytokines due to fibroblasts, fibrocytes and some leukocytes in its content.</p></div>","PeriodicalId":11675,"journal":{"name":"Ejc Supplements","volume":"13 1","pages":"Page 30"},"PeriodicalIF":0.0,"publicationDate":"2015-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.ejcsup.2015.08.053","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54309560","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"P56","authors":"D. Meshalkina,&nbsp;M. Shevtsov,&nbsp;B. Margulis,&nbsp;I. Guzhova","doi":"10.1016/j.ejcsup.2015.08.065","DOIUrl":"10.1016/j.ejcsup.2015.08.065","url":null,"abstract":"<div><p>Glioblastoma is one of the most malignant cancer types. Its median survival is 15<!--> <!-->months with combined radio- and chemotherapy and only 4<!--> <!-->months without therapy. Molecular chaperones play a very multifaced role in tumor development. Depletion of Hdj1 in cancer cells accelerates tumor growth. Decrease of Hdj2 level correlates with the increase of tumor aggressiveness, but also attenuates tumor protection against radiotherapy. Hsp70 provides considerable survival advantages to the cancer cells, but at the same time can act as a “chaperokine”, activating antitumoral immunity. For assessment of chaperone’s role in glioma progression, invasiveness and metastasis formation we chose rat model of intracranial injection of 10⁵ C6 cells. We developed three C6-based cell lines with protein knock-down by RNA-interference: C6 shHsp70 (on 83%), C6 shHdj1 (on 96%) and C6 shHdj2 (on 52%). The last differed in roundish and easily detachable morphology. Following intracranial injection of the modified tumor cells, the animals’ survival was estimated. As compared to the groups of control C6 (25.4<!--> <!-->±<!--> <!-->3.9<!--> <!-->days) and C6 shHdj1 (25.5<!--> <!-->±<!--> <!-->3.8) we observed a nearly 1.5-fold decrease in survival in C6 shHdj2 (16.8<!--> <!-->±<!--> <!-->3.5<!--> <!-->days) (<em>P</em> <!-->&lt;<!--> <!-->0.05). On the contrary, in C6 shHsp70 group the survival increased up to 42.5<!--> <!-->±<!--> <!-->12.0<!--> <!-->days (the increase is completely explainable by the slower growth rate of the culture). Subsequent MR imaging and histological analysis of tumors demonstrated elevated invasiveness and metastatic activity in C6 shHdj2 group in comparison to C6 shHsp70, C6 shHdj1 and control C6. High migration activity and the ability of floating C6 shHdj2 cells to adhere and settle on the substrate was proved in wound-healing assay, spot-healing assay, colony forming assay and transwell migration assay. Adhesion assay showed decreased adhesion ability of C6 shHdj2 cells and increased – of C6 shHsp70 cells on all types of tested extracellular matrixes. Immunofluorescence analyses showed loss of membrane- expressed N-cadherin and loss of intercellular contacts mediated by N-cadherin in C6 shHdj2 cells in comparison to other considered cell lines (although its level in western blot was elevated). Actin staining with rhodamine-falloidin revealed highly abundant leading edges in C6 shHdj2 culture. Matrix metalloprotease zymography proved an increased activity in gelatinases (mmp2 and mmp9) as well as in caseinases (mmp1 and mmp8) in C6 shHdj2 culture supernatant. Assay of stemness marker CD133 expression showed its 11.8 times increase in C6 shHdj2.</p><p>Our experiments proved the high importance of Hdj2 level in glioma progression, invasion and metastasis.</p></div>","PeriodicalId":11675,"journal":{"name":"Ejc Supplements","volume":"13 1","pages":"Pages 36-37"},"PeriodicalIF":0.0,"publicationDate":"2015-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.ejcsup.2015.08.065","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54309742","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"T26","authors":"Z. Pranjol ,&nbsp;N. Gutowski ,&nbsp;M. Hannemann ,&nbsp;J. Whatmore","doi":"10.1016/j.ejcsup.2015.08.078","DOIUrl":"10.1016/j.ejcsup.2015.08.078","url":null,"abstract":"<div><h3>Background</h3><p>Epithelial ovarian cancer frequently metastasizes to the omentum, a process that requires pro-angiogenic activation of HOMECs by tumour -secreted factors in their microenvironment. We have previously shown that ovarian cancer cells secrete a range of factors with possible roles in metastatic angiogenesis including the lysosomal proteases cathepsin D (CD) and cathepsin L (CL). However, the role of these proteases in ovarian cancer metastasis to the omentum is not fully understood.</p></div><div><h3>Aim</h3><p>To investigate whether proliferative effects of CD and CL are dependent on their catalytic activity in HOMECs.</p><p>To investigate the intracellular signalling kinases activated by CD and CL.</p></div><div><h3>Method</h3><p>HOMEC proliferation was assessed by using a colorimetric (WST1) assay. Potential signalling pathways were examined by phosphokinase array and ELISAs. pH experiments were carried out examine whether the observed effects were due to the catalytic activity of CD and CL.</p></div><div><h3>Result</h3><p>CD and CL (50<!--> <!-->ng/ml) significantly increased HOMEC proliferation to 141<!--> <!-->±<!--> <!-->27% (<em>p</em> <!-->=<!--> <!-->0.001, <em>n</em> <!-->=<!--> <!-->50) and 151%<!--> <!-->±<!--> <!-->34% (<em>p</em> <!-->=<!--> <!-->0.001, <em>n</em> <!-->=<!--> <!-->45) respectively vs. control (100%) 72<!--> <!-->h post-treatment. Inhibitors of CD and CL enzyme activity had no effect on HOMEC proliferation and subsequent pH data suggest a nonproteolytic mitogenic activity of these cathepsins. Both proteins induced phosphorylation of ERK1/2, AKT and p38<em>α</em> to ∼2, ∼1.5 and ∼1.5 folds respectively relative to total levels (compared to control).</p></div><div><h3>Conclusion</h3><p>CD and CL induced proliferation in HOMECs. CD and CL may non- proteolytically contribute to pro-angiogenic responses of the omental microvasculature. Induction of phosphorylation of proliferative kinases ERK1/2, AKT and p38<em>α</em> suggest possible downstream signalling cascades of these proteins.</p></div>","PeriodicalId":11675,"journal":{"name":"Ejc Supplements","volume":"13 1","pages":"Page 44"},"PeriodicalIF":0.0,"publicationDate":"2015-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.ejcsup.2015.08.078","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54309913","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"P90","authors":"A. Ponomaryova ,&nbsp;E. Rykova ,&nbsp;N. Cherdyntseva ,&nbsp;E. Morozkin ,&nbsp;I. Zaporozhchenko ,&nbsp;T. Skvortsova ,&nbsp;A. Dobrodeev ,&nbsp;A. Zav’yalov ,&nbsp;S. Tuzikov ,&nbsp;V. Vlassov ,&nbsp;P. Laktionov","doi":"10.1016/j.ejcsup.2015.08.077","DOIUrl":"10.1016/j.ejcsup.2015.08.077","url":null,"abstract":"&lt;div&gt;&lt;h3&gt;Background&lt;/h3&gt;&lt;p&gt;The analysis of circulating tumor nucleic acids (DNAs and RNAs) in the blood seems to be a promising approach for the development of the low-invasive methods of tumor detection, valuable for clinical practice. Detection of oncogenic and tumor suppressor miRNAs in the blood plasma/serum is evidence of their participation in pathogenesis and suggest the possibility of their use as tumor markers and targets for therapy. Thus, the determination of expression level of tumor-associated miRNAs in plasma can lead to significant progress in understanding the tumor development and treatment.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Aim&lt;/h3&gt;&lt;p&gt;Estimation the changes in expression level of miRNAs (miR-19b, miR-25, miR-125b, miR-126, miR-205) in blood plasma from lung cancer patients during combined therapy and to estimate their value as disease monitoring markers.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Materials and methods&lt;/h3&gt;&lt;p&gt;Blood samples were taken from patients (n&lt;!--&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;23) with non-small cell lung cancer treated at the Tomsk Cancer Research Institute. These samples were stabilized and fractionated into plasma and blood cells. MicroRNA was isolated from blood plasma using single-phase phenol-free extraction protocol and purified on silica-based spin columns (BioSilica Ltd, Novosibirsk, Russia). Concentration of five above mentioned miRNAs was measured by quantitative RT-PCR and normalized to miR-16 using dCt method.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Results&lt;/h3&gt;&lt;p&gt;In this study we analyzed the dynamic expression changes of circulating DNA in blood plasma from lung cancer patients during the combined therapy. Circulating miRNAs were isolated from plasma samples of non-small cell lung cancer patients before treatment, within 30&lt;!--&gt; &lt;!--&gt;days after completing chemotherapy and 15&lt;!--&gt; &lt;!--&gt;days after surgery, by using developed methodological approach. In case of miR-19b and miR-125b analysis was found that the miRNA expression level correlates with clinical response to chemotherapy and surgery. Increasing level of miR-19b and decreasing level of miR-125b were associated with therapeutic response. Using Repeated measures ANOVA analysis we demonstrated that the miR-19b and miR-125b expression levels changes throughout three check-up points during the combined treatment are characterized by the significant cubic trend (&lt;em&gt;P&lt;/em&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;0.00284 and &lt;em&gt;P&lt;/em&gt; &lt;!--&gt;=&lt;!--&gt; &lt;!--&gt;0.029, respectively). Changes of miRNA-126 expression level during post-treatment follow-up were not characterized by the definite trend and not correlated with changes of other miRNAs expression level. It was shown the significant correlation between miRNA-25 and miRNA-205 expression levels, but was not found the trend of these miRNAs level changes.&lt;/p&gt;&lt;/div&gt;&lt;div&gt;&lt;h3&gt;Conclusion&lt;/h3&gt;&lt;p&gt;The clinical utility of the circulating miR-19b and miR-125b expression analysis from this study remains to be validated in large cohorts of patients with different histological types of tumors, at the differ","PeriodicalId":11675,"journal":{"name":"Ejc Supplements","volume":"13 1","pages":"Pages 43-44"},"PeriodicalIF":0.0,"publicationDate":"2015-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.ejcsup.2015.08.077","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54309900","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"P108","authors":"E. Rybalkina ,&nbsp;G. Pavlova ,&nbsp;N. Moiseeva ,&nbsp;O. Susova ,&nbsp;A. Mitrofanov ,&nbsp;D. Panteleev ,&nbsp;N. Pustogarov","doi":"10.1016/j.ejcsup.2015.08.085","DOIUrl":"10.1016/j.ejcsup.2015.08.085","url":null,"abstract":"<div><h3>Background</h3><p>Glioblastomas (GBL) are most aggressive brain tumors in adults. They are often radio- and chemotherapy resistant. The standard therapy of GBL includes surgery followed by radiotherapy with concomitant chemotherapy with temozolomide named temodal (imidazotetrazine derivative). Methylation of the O6-methylguanine-DNA-methyltransferase (MGMT) gene promoter is considered as predictor of better outcome of temodal therapy in comparison with the cases without MGMT promoter methylation (Stupp et al., 2009). The aim of this study is finding of additional predictors of temozolomide therapy effectiveness.</p></div><div><h3>Materials and methods</h3><p>We obtained primary cultures of GBL. The rate of GBL sensitivity to temozolomide was revealed by MTT test. Expression of multidrug resistance genes and genes which mediate temozolomide sensitivity (YB1, MDR1, MRP1, LRP, BCRP and MGMT) was determined by real time PCR. Protein localization and expression was revealed by immunohistochemical technique.</p></div><div><h3>Results</h3><p>We developed 14 primary GBL cultures. Staining for nestin was used for demonstration of neuronal origin of cell cultures. Analysis of gene expression and the rate of cell proliferation of the cultures showed that the rate of YB1 gene expression in slow proliferating cultures is significantly lower than in quickly proliferating cell populations. Cell cultures differed in temodal sensitivity 2–2.5-fold. We analyzed whether GBL cell sensitivity to temodal and the rate of several genes’ expression are correlated. We did not find the correlation between cell temodal sensitivity and the rate of MGMT gene expression. These data are in accordance with some results of another authors (Mullins et al., 2013; Brennan et al., 2013). However, we revealed positive correlation of the temodal sensitivity of GBL cultures and MVP/LRP expression (<em>r</em> <!-->=<!--> <!-->0.5592, <em>p</em> <!-->=<!--> <!-->0.04). We found also the trend to negative correlation between GBL temodal sensitivity and YB1 gene expression (<em>r</em> <!-->=<!--> <!-->−0.43602, <em>p</em> <!-->=<!--> <!-->0.02) as well as MDR1 expression (<em>r</em> <!-->=<!--> <!-->−0.4195, <em>p</em> <!-->=<!--> <!-->0.02).</p></div><div><h3>Conclusion</h3><p>The rate of temodal sensitivity of GBL primary cultures is connected with the rate of the expression of MVP/LRP, YB1 and MDR1genes. It is likely that YB-1 protein affects temodal sensitivity by influence on DNA reparation. YB1 could also have an effect on MDR1 expression. MVP/LRP expression is independent factor of GBL prognosis.</p><p><em>This paper was completed with partial support from the</em> <span><em>Russian Foundation for Basic Research</em></span> <em>– Russia (Grant No.</em> <span><em>23-04-40204</em></span><em>).</em></p></div>","PeriodicalId":11675,"journal":{"name":"Ejc Supplements","volume":"13 1","pages":"Pages 47-48"},"PeriodicalIF":0.0,"publicationDate":"2015-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.ejcsup.2015.08.085","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54310002","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"P28","authors":"M. Aksenenko,&nbsp;T. Ruksha","doi":"10.1016/j.ejcsup.2015.08.002","DOIUrl":"10.1016/j.ejcsup.2015.08.002","url":null,"abstract":"&lt;div&gt;&lt;p&gt;Melanoma is one of the aggressive cancer types. Mutations that lock the BRAF protein in an active state may cause excessive signaling in the pathway, leading to uncontrolled cell growth and survival. Primarily among the BRAF mutations observed in melanoma, over 90% are supposed to be at codon 600, resulting in substitution of glutamic acid for valine, V600E (T&lt;!--&gt; &lt;!--&gt;&gt;&lt;!--&gt; &lt;!--&gt;A transversion) located in exon 15, BRAFV600E. Of particular interest is BRAF negative melanoma. This type of melanoma is not sensitive to BRAF inhibitors and approaches to therapy require further study. We aimed to investigate the frequency of BRAF V600 mutations in 80 patients with primary melanoma and determine the relationship between mutations and clinical/pathologic features. Genomic DNA was extracted from biopsy specimens with prevalent percentage of tumor cells by DNA-sorb B isolation kit (Amplisense, Russia). BRAF V600E mutation was estimated by real-time PCR-based assay for the BRAF V600E mutation allele-specific DNA test (BioLink, Russia). Breslow thickness was assessed by applying commercial Infinity Capture, Infinity Analyze Software. The lymphocytic infiltration was determined in all tumors and classified as “brisk”, “nonbrisk”, and “absent” according to criteria established by Clark et al. Tumor infiltrating lymphocytes (TIL) were identified as lymphocytes within tumor nodes. “Brisk” infiltrate was determined in case of a diffuse presence of lymphocytes within tumor, “non-brisk” infiltrate was in focal location of lymphocytes and “absent” if no lymphocytes were present in a tumor. Mitotic activity was determined as mitotic count on 10 high power fields. For all patients, clinical and pathologic features were tested for significant association with BRAF V600E mutation status using simple cross tabulations, Fisher’s exact test, Pearson’s &lt;em&gt;χ&lt;/em&gt;&lt;sup&gt;2&lt;/sup&gt; test, and/or non-parametric Mann–Whitney &lt;em&gt;U&lt;/em&gt; test. The &lt;em&gt;P&lt;/em&gt; values lower 0.05 were considered as significant. BRAF V600E mutation was detected in 41.25% of tested tumours. Patients with BRAF-mutant and non-BRAF mutant melanoma were matched by age and gender. Superficial spreading melanomawas observed in 66.2% of patients with wild-type BRAF, nodular melanoma in 21.2%, both lentigo-melanoma and acral-lentiginous melanomain 6.3% and mucosal melanoma in 1 3.0% of patients. In wild-type BRAF melanoma patients, 59.7% tumors had “brisk” infiltrate, 14.8% – “non-brisk”, and 12 25.5% had no infiltrate. There was no found correlation between BRAF status and tumor localization, clinico-pathological type of tumor, TIL status, Breslow thickness and mitotic rate. However, when cases were stratified by age, it was revealed that melanoma patients aged above 80&lt;!--&gt; &lt;!--&gt;years were preferentially BRAF-negative (&lt;em&gt;p&lt;/em&gt; &lt;!--&gt;&lt;&lt;!--&gt; &lt;!--&gt;0.05). BRAF-negative melanomas occurred significantly more frequent in superficial spreading type of the tumor. The localization of melanomas was differe","PeriodicalId":11675,"journal":{"name":"Ejc Supplements","volume":"13 1","pages":"Page 1"},"PeriodicalIF":0.0,"publicationDate":"2015-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.ejcsup.2015.08.002","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54308582","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}