Ejc Supplements最新文献_第9页

P156 P156

Ejc Supplements Pub Date : 2015-11-01 Epub Date: 2015-11-23 DOI: 10.1016/j.ejcsup.2015.08.048

D. Kokova , N. Dementeva , N. Cherdyntseva , A. Gratchev , Julia Kzhyshkowska

{"title":"P156","authors":"D. Kokova , N. Dementeva , N. Cherdyntseva , A. Gratchev , Julia Kzhyshkowska","doi":"10.1016/j.ejcsup.2015.08.048","DOIUrl":"10.1016/j.ejcsup.2015.08.048","url":null,"abstract":"<div><h3>Background</h3><p>Identification of DNA-based biomarkers of cancer cells is highly promising and rapidly developing direction that can advance early detection and therapy of malignancies. DNA adducts are felicitous markers of cancer, because their chemical structure is significantly different from that of mutated or methylated DNA, that allows to determine them with high precision using mass spectrometry. The aim of this work is to develop the methodology of sample preparation and its mass spectrometric analysis. Samples were prepared from the blood plasma and from the tumor tissue from lung cancer patients and from blood of healthy individuals.</p></div><div><h3>Materials and methods</h3><p>DNA was isolated from the blood plasma and tissue by using column method (BioSilica, Russia) The final yield from 1ml of blood was 100ng. DNA samples were subjected to acid hydrolysis (1M HCl) at 70°C. After 3h, the hydrolysis was stopped by cooling on ice for 5min and later on adding an equivalent amount of an alkali and a phosphate buffer solution (pH 7). To assess the extent of hydrolysis of the samples they were analysed by electrophoresis on a 1.2% agarose gel in Tris-acetate buffer. The samples were extracted at cartridge HF Bond Elut-C18 100mg, 1ml (Agilent Technologies, USA) and eluted in several fractions with a gradual increase of methanol in the eluent. Stream of nitrogen was applied to dry the extract. The samples were subjected to mass spectrometric analysis after pre-separation by UHLC Ultimate 3000 RS (Dionex, USA) in a column Dionex Acclaim RSLC 120 C18 (2.1×50mm 120A, 0.2μm) flow rate of 0.5ml/min using as eluents 0.1% solution of formic acid in water (A) and 0.1% solution of formic acid in atsetontrile (B). Elution was carried out in gradient mode: (%B): 0–3min (5%), 3–28min (5–95%), 28–30min (95%), 30–31min (95–5%), 31–35min (5%). Mass spectrometry was carried out on ESI-qTOF ultrahigh resolution Maxis 4G (Bruker, Germany) in the positive ion detection mode range 50–1000m/z, 2Hz with the following settings electrospray ion source: CV 3800V, Nebulizer gas 1bar, Dry Gas 8 l/min, Dry Temp: 200°C.</p></div><div><h3>Results</h3><p>It was found that the DNA which was cleaved with acid hydrolysis in the result contained single DNA bases. The samples were stable at 4°C for at least 7days. The optimal eluent for solid phase extraction of DNA is 80% solution of methanol in water. The number of DNA adducts was evaluated by the integrated value of the mass spectrometric response detector. It was shown that most amount of adducts 4-hydroxy-1-(3-pyridyl)-1- butanone and N3-(2-carbamoyl-2-hydroxyethyl)adenine was found in DNA samples derived from tumor tissue. The adduct N7-(2-carbamoyl-","PeriodicalId":11675,"journal":{"name":"Ejc Supplements","volume":"13 1","pages":"Pages 27-28"},"PeriodicalIF":0.0,"publicationDate":"2015-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.ejcsup.2015.08.048","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54309493","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}

引用次数: 1

P71 P71

Ejc Supplements Pub Date : 2015-11-01 Epub Date: 2015-11-23 DOI: 10.1016/j.ejcsup.2015.08.058

A. Leshchenko , N. Matsenko

{"title":"P71","authors":"A. Leshchenko , N. Matsenko","doi":"10.1016/j.ejcsup.2015.08.058","DOIUrl":"10.1016/j.ejcsup.2015.08.058","url":null,"abstract":"<div><h3>Background</h3><p>Breast cancer is the most common cause of death from cancer among women aged 40–69years. According to WHO, about 1 million new cases of breast cancer are diagnosed annually worldwide, and nearly 500,000 die from it. In Russia, more than 55,000 women are diagnosed with breast cancer annually, and more than 22,000 die from it. Diagnosis and treatment of breast cancer is the primary and important social and medical problem.The increased expression of: (a) ER and PgR (70–75% of all cases of breast cancer) is an indication for hormone therapy, which is one of the simplest and most effective methods of systemic treatment of breast cancer; (b) receptor HER2/neu is a marker of highly aggressive form of breast cancer and indication for the use of targeted therapy Gertseptin; (c) proliferative factor Ki-67 reflects the ability of a tumor to metastasize.</p><p>Today, the “gold standard” of gene expression diagnostic of ER, PgR, HER2/neu and Ki-67 in breast cancer is immunohistochemistry (IHC) using foreign test systems (Ventana, Dako Inc, USA) . However, IHC diagnostics has some significant drawbacks. It leads up to 15–17% of cases of incorrect choice of drug therapy, which based on incorrect results of IHC studies. As a result, the significant group of patients do not receive effective treatment. Aim of the study Aim: the development a prototype of diagnostic test system for detection the receptor status of breast cancer, based on RT-PCR.</p></div><div><h3>Materials and methods</h3><p>Breast cancersamples consisted of 45 fresh-frozen tissue (FFT) samples of breast cancer (sites of malignant transformation and normal tissue from the same patients) and 59 FFPET sampleswere collected. All the samples had the IHC characteristic of receptor status of ER, PgR, HER2/neu and proliferation factor Ki-67 (Dako Inc., USA). Samples were submitted by SBIH NR “Novosibirsk Regional Oncology Center” (Novosibirsk, Russia). The experimental part of the study was divided into several stages: separation of mRNA from cells, obtaining cDNA (reverse transcription reaction), PCR in real-time and validation of the test system. Isolation of total RNA from FFT samples was performed using a set of “SV Total RNA Isolation system” according to the manufacturer’s instructions (Promega, USA). Isolation of total RNA from FFPET samples was performed using a set “ReliaPrep FFPE Total RNA Miniprep System” (Promega, USA) according to the manufacturer’s instructions. The concentration of total RNA was determined using a NanoDrop 1000 microspectrophotometer (Thermo Bioscience, USA) (RNA concentration were 15–660ng/ml).</p></div><div><h3>Results</h3><p>For the reverse transcription reaction (RT) and PCR, the main parameters were selected. For RT: RNA incubation time and temperature of the reaction, enzyme concentration in the reaction mix. For PCR: the amount of DNA template, the number of primers, the concentration of magny ions, the concentration","PeriodicalId":11675,"journal":{"name":"Ejc Supplements","volume":"13 1","pages":"Pages 32-33"},"PeriodicalIF":0.0,"publicationDate":"2015-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.ejcsup.2015.08.058","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54309629","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}

引用次数: 0

T35 T35

Ejc Supplements Pub Date : 2015-11-01 Epub Date: 2015-11-23 DOI: 10.1016/j.ejcsup.2015.08.044

F. Kisseljov, S. Vinokurova, N. Kisseljova, L. Pavlova, M. Fedorova, A. Katargin, A. Petrenko, L. Korolenkova

{"title":"T35","authors":"F. Kisseljov, S. Vinokurova, N. Kisseljova, L. Pavlova, M. Fedorova, A. Katargin, A. Petrenko, L. Korolenkova","doi":"10.1016/j.ejcsup.2015.08.044","DOIUrl":"10.1016/j.ejcsup.2015.08.044","url":null,"abstract":"<div><p>Epigenetics investigates mechanisms that control inheritance of gene expression program during somatic cell divisions. These mechanisms include regulation by DNA methylation, histone post-translational modifications and nucleosome positioning, functioning of regulatory non-coding RNAs, control of alternative splicing of mRNA precursors and high-order chromatin organization. Genome-wide loss of epigenetic stability and increased epigenetic plasticity are common features of all tumor types. In normal tissues epigenetic plasticity allow cells to response on environment signals. Thus, in tumor cells its constitutive activation leads to epigenetic heterogeneity that are the additional hallmark of the most of the classical cancers. Cervical cancers are one of the most interesting models for the analysis of the role of epigenetic changes in tumor progression. These types of tumors are associated with infection of human papilloma viruses of so-called high-risk group (HR-HPV) and characterized by well-defined stages of malignant conversion from intraepithelial neoplasias to carcinomas. The viral DNA can persist in episomal form or integrates into the host-cell genome.</p><p>Cellular genomes encode genetic information in their linear sequence, but appropriate gene expression requires chromosomes to fold into dynamic complex three-dimensional structures. Scaffold/matrix attachment regions (S/MARs) are specialized genomic DNA sequences that take part in organization of these structures. We demonstrated that methylation of S/MARs was required for their attachment to nuclear matrix and that methylation status of S/MARs was changed in cervical cancer cell compared to normal cells.</p><p>DNA methylation plays an important role in the regulation of gene expression. We found that methylation of the regulatory sequences in the HPV16 genome specifically changes in transformed compared to the normal cervical epithelial cells. Next, we showed that methylation of the transcription factor binding sites modulates the viral oncogene expression. These data suggest that the HPV16 genome methylation may represent an important mechanism that initiates the development of HPV-associated tumors.</p><p>Using next generation sequencing, we identified pattern of differentially expressed microRNAs in clinical samples of the cervical lesions. We confirmed expression of microRNAs that have been described previously as well as identified new microRNAs that can be potentially involved in the development and progression of cervical cancer. Spectrum of differentially expressed microRNAs includes microRNAs targeting tumor-suppressor genes as well as oncogenes.</p><p>Telomerase is a key regulator of cell proliferation. This enzyme is silent in normal cells and activated in most of the tumors. Few forms of RNA (hTERT), encoded by telomerase gene were detected in different tumor cells and among them three forms (alfa, beta and gamma) are most well pronounced. We found that in cervical","PeriodicalId":11675,"journal":{"name":"Ejc Supplements","volume":"13 1","pages":"Page 25"},"PeriodicalIF":0.0,"publicationDate":"2015-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.ejcsup.2015.08.044","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54309431","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}

引用次数: 0

A84: Polymorphic markers Arg72Pro and Gln157Lys of TP53 gene in nonsmall cell lung cancer A84:非小细胞肺癌TP53基因多态性标记Arg72Pro和Gln157Lys

Ejc Supplements Pub Date : 2015-11-01 DOI: 10.1016/j.ejcsup.2015.08.122

T. Zavarykina, A. Byrdennyy, V. Loginov, M. V. Atkarskaya, Zhizhina Gp

引用次数: 0

P76: Mechanics–mathematical models of changing of human cells properties under the cancer action P76:力学——癌症作用下人体细胞特性变化的数学模型

Ejc Supplements Pub Date : 2015-11-01 DOI: 10.1016/J.EJCSUP.2015.08.128

M. Zhuravkov, N. Romanova

引用次数: 0

A14: Heterogeneity in breast tumor microenvironment: A report from one case A14:乳腺肿瘤微环境的异质性:1例报告

Ejc Supplements Pub Date : 2015-11-01 DOI: 10.1016/J.EJCSUP.2015.08.109

L. Tashireva, E. Denisov, O. Savelieva, M. Zavyalova, E. Kaigorodova, E. Slonimskaya, V. Perelmuter

引用次数: 0

P52: Glioma-derived soluble factors pose strong chemoattractants and partially change the cytotoxic activity of IFN-alpha-induced dendritic cells P52:胶质瘤衍生的可溶性因子具有很强的化学引诱剂，并部分改变ifn - α诱导的树突状细胞的细胞毒性活性

Ejc Supplements Pub Date : 2015-11-01 DOI: 10.1016/j.ejcsup.2015.08.114

T. Tyrinova, O. Leplina, M. Tikhonova, S. Mishinov, S. Chernov, Sakhno Lv, V. Stupak, E. Chernykh

引用次数: 0

T125 T125

Ejc Supplements Pub Date : 2015-11-01 Epub Date: 2015-11-23 DOI: 10.1016/j.ejcsup.2015.08.005

O. Bajenova, I. Evsyukov, S. O’Brien

{"title":"T125","authors":"O. Bajenova, I. Evsyukov, S. O’Brien","doi":"10.1016/j.ejcsup.2015.08.005","DOIUrl":"10.1016/j.ejcsup.2015.08.005","url":null,"abstract":"<div><p>Tumor markers play an important role in the identification of human malignancies. It has been shown that the carcinoembryonic antigen (CEA, CEACAM5) is a promoter of metastasis in epithelial cancers that is widely used as a clinical marker. The aim of this study is to elucidate the network of genes that are involved in the CEA-induced liver metastasis. Previously, we have shown that CEA is accumulated in the lungs and livers of rats by interacting with their macrophages. We identified and cloned a new gene (CEAR) for the CEA-binding protein, which is located on the surface of fixed liver macrophages, Kupffer cells (Bajenova et al, 2001). It has been shown that the interaction of CEA and CEAR proteins increases the production of IL-1, IL-10, IL-6, TNF-<em>α</em> cytokines (Thomas et al, 2011). This interaction changes the expression of liver adhesion molecules that enhances the survival of cancer cells to the liver. We also suggested that CEA synthesis by cancer cells may influence the E-cadherin adhesion junction complexes and have shown that CEA production violates the functional relationship between Ecadherin and its partners <em>α</em>-, <em>β</em>- and p120 catenin. A new type of interaction was discovered between the CEA and <em>β</em>-catenin and the increased amount of <em>β</em>-catenin in the nuclei of CEA producing cells. The data show that CEA production can cause the dissociation of cancer cells and trigger cancer progression. The CEA synthesis also alters splicing of p120 catenin protein and causes the release of soluble E-cadherin. Previously, CEA and epithelial E-cadherin were considered as independent tumor markers. Our data explain the correlation between the elevated levels of CEA and the increase in soluble E-cadherin in the progression of colorectal cancer (Bajenova et al, 2014).</p><p>We carried out a comparative transcriptome analysis of CEA-producing cell lines. The RNA transcriptome libraries were obtained and sequenced. By pairwise comparisons of CEA producing and non-producing cell lines using Cummerband program, we selected the set of genes (90 total genes) whose expression have been changed in the CEA-producing cell lines (overexpressed or downregulated). The biological processes that are linked to this differential gene expression were identified by Gene Set Enrichment Analysis (GSEA). In total, 8 significantly enriched GO terms related to the cellular components and biological processes were identified. Using KEGG and GO databases, we also identified the signaling pathways involved in the response to CEA. These findings have direct medical application, since they allow not only to establish the relationships between the existing biomarkers but also to discover the new ones. These biomarkers can be used for diagnosis and monitoring of metastatic carcinomas and for the drug development.</p></div>","PeriodicalId":11675,"journal":{"name":"Ejc Supplements","volume":"13 1","pages":"Page 3"},"PeriodicalIF":0.0,"publicationDate":"2015-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.ejcsup.2015.08.005","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54308649","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}

引用次数: 0

P131 P131

Ejc Supplements Pub Date : 2015-11-01 Epub Date: 2015-11-23 DOI: 10.1016/j.ejcsup.2015.08.016

V. Bychkov, E. Nikitina, N. Litviakov

{"title":"P131","authors":"V. Bychkov, E. Nikitina, N. Litviakov","doi":"10.1016/j.ejcsup.2015.08.016","DOIUrl":"10.1016/j.ejcsup.2015.08.016","url":null,"abstract":"<div><h3>Background</h3><p>The aim of the study was to identify the factors that determine outcome and overall survival of HNC patients of Tomsk region.</p></div><div><h3>Materials and methods</h3><p>Clinical data, morphological characteristics of tumors and outcomes were obtained for 91 patients. Data about lifestyle, food preferences, smoking history were obtained from the questionnaire (<em>n</em>  =0.03) as well as regional lymph nodes status (<em>p</em>  =0.09, <em>p</em>  =0.09, respectively). Overall survival of patients with lymph node metastasis was higher in case of early tumor stages (<em>p</em>  =0.06). Cox regression analysis was used to obtain the model describing overall survival of patients. The model with the highest level of significance includes 3 factors-nodal metastases, the presence of keratinization and radiotherapy. It was shown that the risk of death was 4.2 and 2.6-fold higher in case of lymph node metastases and keratinized cancer, and 2.7-fold lower in case of radiotherapy. It was also shown that metastasis occurred more frequently in cases with invasion into the underlying tissue of a primary tumor (<em>p</em>  =0.02). Association of alcohol consumption with questionnaire data was studied. It was shown that men’s preferably smokers consume alcohol more often than other patients (<em>p</em>  =0.02, respectively). Our data showed that HPV prevalence was higher in smokers (<em>p</em>  =0.07). Furthermore, response to radiotherapy was better in","PeriodicalId":11675,"journal":{"name":"Ejc Supplements","volume":"13 1","pages":"Pages 9-10"},"PeriodicalIF":0.0,"publicationDate":"2015-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.ejcsup.2015.08.016","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54308804","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}

引用次数: 0

A129 A129

Ejc Supplements Pub Date : 2015-11-01 Epub Date: 2015-11-23 DOI: 10.1016/j.ejcsup.2015.08.020

N. Dashenkova, A. Mikaelyan

{"title":"A129","authors":"N. Dashenkova, A. Mikaelyan","doi":"10.1016/j.ejcsup.2015.08.020","DOIUrl":"10.1016/j.ejcsup.2015.08.020","url":null,"abstract":"<div><p>IGF signaling pathway plays an important role in the regulation of cell growth, proliferation, differentiation, apoptosis and survival. Deregulation of this pathway has been frequently identified in the development of hepatocellular carcinoma (HCC). IGF signaling pathway consists of IGF ligands (IGF-I and IGF-II), IGF binding proteins (IGFBP 1-7), and membrane-bound IGF receptors (IGF-1R, IGF-II/M6PR, and IGF-2R). The insulin-like growth factor binding proteins (IGFBPs) have several functions, such as IGF transporting, accumulation of IGF at the specific cell pools, inhibition and activation of ligand–receptor interaction. Furthermore, the Igfbps may act independently of IGF-pathway.</p><p>Diethylnitrosamine (DEN) was used for induction of hepatic tumorigenesis. The male mice, 2weeks old, were injectedintraperitoneally with 20mg/kg body weight of DEN. The liver tissue samples were collected 4, 6, 8, 12months after injection. We performed a comprehensive histological analysis of all kinds of liver samples: tumors, tumor surrounding tissue and normal tissue. Dysplasia was observed 4months after injection, at hepatocellular adenomas were identified 6–8months after injection. In the experimental group, 100% of mice had multifocal HCC 12months after injection. Evaluation of samples from the control group showed normal liver architecture and histology.</p><p>Western blot analysis showed the presence the HCC tumor marker alfa-fetoprotein only in tumor samples. High levels of Igf-1R and FoxO3 proteins were observed in tumor tissue in contrast to the surrounding and normal tissues 12months after injection. The expression profiles of Igfbp-1,-2,-3,-4,-5,-6,-7, Igf-1R, FoxO3 were analyzed by real time PCR. The most significant results were observed 8 and 12months after DEN injection. RT-PCR showed the dynamic increase in Igf-1R expression in tumor. Expression pattern of Igfbps has shown a high level of Igf-1,-2,-5,-6,-7 mRNA in surrounding tissue and Igfbp-1,-3,-5 in tumor compared to control. The expression of Igfbp-2,-4,-5,-6,-7 was up-regulated, whereas expression of Igfbp-1,-3 was down-regulated in surrounding tissue. The expression of Igfbp-4 was decreased in tumor samples. The expression levels of Igfbp-3,-4 in surrounding tissue and Igfbp-7 in tumor were the same as in the control. We detected high level of mRNA of FoxO3 in tumor.</p><p>In conclusion, these data confirm previously existing assumption about paracrine effect of Igfbps on tumor growth and progression. Phosphorylation of FoxO3 protein may play an important role in survival of cancer cells. Our results showed up-regulation of Igfbp-2 and Igfbp-5 in tumor and tumor surrounding tissue. Igfbp-2 and Igfbp-5, well known activators of Igfs, can be pro-tumorigenic factors, which are important for hepatocarcinogenesis. Hence, therapeutic targeting of these proteins may offer options for intervention in","PeriodicalId":11675,"journal":{"name":"Ejc Supplements","volume":"13 1","pages":"Pages 11-12"},"PeriodicalIF":0.0,"publicationDate":"2015-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.ejcsup.2015.08.020","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54308891","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}

引用次数: 0