Pub Date : 2015-11-01Epub Date: 2015-11-23DOI: 10.1016/j.ejcsup.2015.08.053
T. Kunts , K. Karpukhina , E. Mikhailova , N. Varaksin , A. Autenshlyus
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.
Objective
Comparative evaluation of cytokine-producing function of the invasive ductal carcinoma and fibroadenoma of the breast in vitro.
Materials and methods
Similar biopsies (V = 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-1β, IL-1Ra, TNFα, IL-2, IL-6, IL-8, IL-10, IL-17, IL-18, VEGF and IFNγ in the supernatant of the tumor were measured with enzyme-linked immunosorbent assay (ELISA).
Results
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 IFNγ 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 IFNγ 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 IFNγ leads to an increase in the IFNγ level. In early stages of tumor development, IFNγ 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 IFNγ 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.
Conclusion
Cytokine production in supernatants of invasive ductal carcinoma compared with fibroadenoma of the breast is characterized by increase in IL-17, IL-18 and IFNγ 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.
{"title":"P64","authors":"T. Kunts , K. Karpukhina , E. Mikhailova , N. Varaksin , 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<!--> <!-->mm<sup>3</sup>) 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}
Pub Date : 2015-11-01Epub Date: 2015-11-23DOI: 10.1016/j.ejcsup.2015.08.065
D. Meshalkina, M. Shevtsov, B. Margulis, I. Guzhova
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) (P < 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.
Our experiments proved the high importance of Hdj2 level in glioma progression, invasion and metastasis.
{"title":"P56","authors":"D. Meshalkina, M. Shevtsov, B. Margulis, 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 10<sup>5</sup> 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> <!--><<!--> <!-->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}
Pub Date : 2015-11-01Epub Date: 2015-11-23DOI: 10.1016/j.ejcsup.2015.08.085
E. Rybalkina , G. Pavlova , N. Moiseeva , O. Susova , A. Mitrofanov , D. Panteleev , N. Pustogarov
Background
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.
Materials and methods
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.
Results
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 (r = 0.5592, p = 0.04). We found also the trend to negative correlation between GBL temodal sensitivity and YB1 gene expression (r = −0.43602, p = 0.02) as well as MDR1 expression (r = −0.4195, p = 0.02).
Conclusion
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.
This paper was completed with partial support from theRussian Foundation for Basic Research– Russia (Grant No.23-04-40204).
胶质母细胞瘤(GBL)是成人中最具侵袭性的脑肿瘤。它们通常对放疗和化疗具有耐药性。GBL的标准治疗包括手术后放疗伴替莫唑胺替莫达(咪唑四嗪衍生物)化疗。与没有MGMT启动子甲基化的病例相比，o6 -甲基鸟嘌呤- dna甲基转移酶(MGMT)基因启动子的甲基化被认为是蝶呤治疗效果更好的预测因子(Stupp et al.， 2009)。本研究的目的是寻找替莫唑胺治疗效果的其他预测因素。材料和方法我们获得了GBL原代培养。MTT试验显示GBL对替莫唑胺的敏感性。实时荧光定量PCR检测多药耐药基因及替莫唑胺敏感性相关基因(YB1、MDR1、MRP1、LRP、BCRP、MGMT)的表达情况。免疫组化技术显示蛋白定位和表达。结果共培养了14株GBL原代培养。巢蛋白染色用于证明细胞培养的神经元来源。对培养物的基因表达和细胞增殖率的分析表明，缓慢增殖培养物中YB1基因的表达率明显低于快速增殖细胞群。细胞培养在温度敏感性上有2 - 2.5倍的差异。我们分析了GBL细胞对temodal的敏感性与几个基因的表达率是否相关。我们没有发现细胞温度敏感性与MGMT基因表达率之间的相关性。这些数据与其他作者的一些结果一致(Mullins et al.， 2013;Brennan et al.， 2013)。然而，我们发现GBL培养物的模态敏感性与MVP/LRP表达呈正相关(r = 0.5592, p = 0.04)。我们还发现，GBL模态敏感性与YB1基因表达(r = - 0.43602, p = 0.02)和MDR1表达(r = - 0.4195, p = 0.02)呈负相关趋势。结论GBL原代培养的模态敏感性与MVP/LRP、YB1和mdr1基因的表达率有关。YB-1蛋白可能通过影响DNA修复来影响模态敏感性。YB1也可能影响MDR1的表达。MVP/LRP表达是影响GBL预后的独立因素。本文由俄罗斯基础研究基金会(资助号:23-04-40204)部分资助完成。
{"title":"P108","authors":"E. Rybalkina , G. Pavlova , N. Moiseeva , O. Susova , A. Mitrofanov , D. Panteleev , 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}
Pub Date : 2015-11-01Epub Date: 2015-11-23DOI: 10.1016/j.ejcsup.2015.08.078
Z. Pranjol , N. Gutowski , M. Hannemann , J. Whatmore
Background
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.
Aim
To investigate whether proliferative effects of CD and CL are dependent on their catalytic activity in HOMECs.
To investigate the intracellular signalling kinases activated by CD and CL.
Method
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.
Result
CD and CL (50 ng/ml) significantly increased HOMEC proliferation to 141 ± 27% (p = 0.001, n = 50) and 151% ± 34% (p = 0.001, n = 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 p38α to ∼2, ∼1.5 and ∼1.5 folds respectively relative to total levels (compared to control).
Conclusion
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 p38α suggest possible downstream signalling cascades of these proteins.
上皮性卵巢癌经常转移到网膜，这一过程需要肿瘤分泌因子在其微环境中激活homec的促血管生成。我们之前已经证明卵巢癌细胞分泌一系列可能在转移性血管生成中起作用的因子，包括溶酶体蛋白酶组织蛋白酶D (CD)和组织蛋白酶L (CL)。然而，这些蛋白酶在卵巢癌大网膜转移中的作用尚不完全清楚。目的探讨CD和CL在HOMECs中的增殖作用是否依赖于它们的催化活性。探讨CD和CL对细胞内信号激酶的激活作用。方法采用比色法(WST1)检测细胞增殖。通过磷酸激酶阵列和elisa检测潜在的信号通路。进行了pH实验，以检验所观察到的效果是否由CD和CL的催化活性引起。结果cd和CL (50 ng/ml)处理72 h后，与对照组(100%)相比，分别使HOMEC增殖141±27% (p = 0.001, n = 50)和151%±34% (p = 0.001, n = 45)。CD和CL酶活性抑制剂对HOMEC增殖没有影响，随后的pH数据表明这些组织蛋白酶具有非蛋白溶解性有丝分裂活性。两种蛋白均诱导ERK1/2、AKT和p38α的磷酸化水平相对于总水平分别达到2倍、1.5倍和1.5倍(与对照组相比)。结论cd和CL可诱导HOMECs细胞增殖。CD和CL可能非蛋白性地促进大网膜微血管的促血管生成反应。诱导增殖激酶ERK1/2、AKT和p38α的磷酸化提示这些蛋白可能存在下游信号级联反应。
{"title":"T26","authors":"Z. Pranjol , N. Gutowski , M. Hannemann , 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}
Pub Date : 2015-11-01Epub Date: 2015-11-23DOI: 10.1016/j.ejcsup.2015.08.077
A. Ponomaryova , E. Rykova , N. Cherdyntseva , E. Morozkin , I. Zaporozhchenko , T. Skvortsova , A. Dobrodeev , A. Zav’yalov , S. Tuzikov , V. Vlassov , P. Laktionov
<div><h3>Background</h3><p>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.</p></div><div><h3>Aim</h3><p>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.</p></div><div><h3>Materials and methods</h3><p>Blood samples were taken from patients (n<!--> <!-->=<!--> <!-->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.</p></div><div><h3>Results</h3><p>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<!--> <!-->days after completing chemotherapy and 15<!--> <!-->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 (<em>P</em> <!-->=<!--> <!-->0.00284 and <em>P</em> <!-->=<!--> <!-->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.</p></div><div><h3>Conclusion</h3><p>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
{"title":"P90","authors":"A. Ponomaryova , E. Rykova , N. Cherdyntseva , E. Morozkin , I. Zaporozhchenko , T. Skvortsova , A. Dobrodeev , A. Zav’yalov , S. Tuzikov , V. Vlassov , P. Laktionov","doi":"10.1016/j.ejcsup.2015.08.077","DOIUrl":"10.1016/j.ejcsup.2015.08.077","url":null,"abstract":"<div><h3>Background</h3><p>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.</p></div><div><h3>Aim</h3><p>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.</p></div><div><h3>Materials and methods</h3><p>Blood samples were taken from patients (n<!--> <!-->=<!--> <!-->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.</p></div><div><h3>Results</h3><p>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<!--> <!-->days after completing chemotherapy and 15<!--> <!-->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 (<em>P</em> <!-->=<!--> <!-->0.00284 and <em>P</em> <!-->=<!--> <!-->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.</p></div><div><h3>Conclusion</h3><p>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}
Pub Date : 2015-11-01Epub Date: 2015-11-23DOI: 10.1016/j.ejcsup.2015.08.002
M. Aksenenko, T. Ruksha
<div><p>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<!--> <!-->><!--> <!-->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 <em>χ</em><sup>2</sup> test, and/or non-parametric Mann–Whitney <em>U</em> test. The <em>P</em> 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<!--> <!-->years were preferentially BRAF-negative (<em>p</em> <!--><<!--> <!-->0.05). BRAF-negative melanomas occurred significantly more frequent in superficial spreading type of the tumor. The localization of melanomas was differe
{"title":"P28","authors":"M. Aksenenko, T. Ruksha","doi":"10.1016/j.ejcsup.2015.08.002","DOIUrl":"10.1016/j.ejcsup.2015.08.002","url":null,"abstract":"<div><p>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<!--> <!-->><!--> <!-->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 <em>χ</em><sup>2</sup> test, and/or non-parametric Mann–Whitney <em>U</em> test. The <em>P</em> 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<!--> <!-->years were preferentially BRAF-negative (<em>p</em> <!--><<!--> <!-->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}
Pub Date : 2015-11-01Epub Date: 2015-11-23DOI: 10.1016/j.ejcsup.2015.08.052
M. Kruchinina , A. Starikov , S. Kurilovich , V. Kruchinin , V. Volodin , S. Rykhlitskii , A. Gromov , S. Peltec , S. Shehovtsov , V. Generalov
<div><h3>Background</h3><p>The aim of this work was to assess the potential of the optical methods for studying erythrocytes (Er) and blood serum (BS) of patients with colorectal cancer (CC).</p></div><div><h3>Methods</h3><p>A total of 26 persons (52<!--> <!-->+<!--> <!-->8<!--> <!-->years old) with CC (histologically – adenocarcinoma) in the T1–2 stage (the 1st group consisting of 10 patients) and in the terminal stage T3–4 (the 2nd group involving 16 patients) were examined. The metastases (in the liver area) were detected in 6 patients; the remaining patients had no metastases. The degree of lymph node involvement in most patients was not determined, the ten corresponding N1. The control group consisted of 16 healthy people (50<!--> <!-->+<!--> <!-->6<!--> <!-->years old). Electric and viscoelastic Er parameters were investigated by dielectrophoresis, their membrane structure – by TLC and gas chromatography. The optical properties of BS were studied by the methods of ellipsometry. The reaction of the monoclonal antibody CD 24 with BS antigens of CC patients was studied by spectroscopic ellipsometry close to the conditions of surface plasmon resonance (SPR) (ProteOn XPR36 (BioRad).</p></div><div><h3>Results</h3><p>We observed significant differences in Er parameters, associated with the CC stage. Given in the 2nd group (T3–4) summarized rigidity, viscosity, electrical conductivity, the relative polarizability, indexes of aggregation and destruction were significantly higher than those in the 1st (T1–2) and in the control group (<em>p</em> <!--><<!--> <!-->0.001–0.05). At the same time the patients of the 2nd group had marked disturbances of Er deformability, leading to the development of microcirculatory disorders and tissue hypoxia with the expressed deficit of intracellular macroergs. We observed high levels of cholesterol fraction, oleic, stearic acids, high index of cholesterol/phospholipids (PHL) and low levels of total lipids, easily oxidable PHL, arachidonic acid, omega-3 index in Er membranes in the 2nd group in comparison with those in the 1st group of patients (<em>p</em> <!--><<!--> <!-->0.0001–0.03). Scanning ellipsometry showed marked heterogeneity in thickness and composition, the abundance of discontinuities in thin films of BS of patients in the 2nd group compared to the 1st one (<em>p</em> <!--><<!--> <!-->0.001). Increasing the refractive index in combination with the reduction in film thickness as CC stage was weighting has been observed (<em>p</em> <!--><<!--> <!-->0.01–0,<span><math><mrow><mo>⧹</mo></mrow></math></span>.05). The concentration of the antigens to the CD24 in the BS of patients (obtained by SPR) in the terminal stages of CC was higher than that in the T1–2 (<em>p</em> <!--><<!--> <!-->0.001). We revealed correlations between Er parameters, BS ellipsometry characteristics and biochemical parameters, which reflected the interaction between these components depending on the CC stage.</p></div><div>
{"title":"P41","authors":"M. Kruchinina , A. Starikov , S. Kurilovich , V. Kruchinin , V. Volodin , S. Rykhlitskii , A. Gromov , S. Peltec , S. Shehovtsov , V. Generalov","doi":"10.1016/j.ejcsup.2015.08.052","DOIUrl":"10.1016/j.ejcsup.2015.08.052","url":null,"abstract":"<div><h3>Background</h3><p>The aim of this work was to assess the potential of the optical methods for studying erythrocytes (Er) and blood serum (BS) of patients with colorectal cancer (CC).</p></div><div><h3>Methods</h3><p>A total of 26 persons (52<!--> <!-->+<!--> <!-->8<!--> <!-->years old) with CC (histologically – adenocarcinoma) in the T1–2 stage (the 1st group consisting of 10 patients) and in the terminal stage T3–4 (the 2nd group involving 16 patients) were examined. The metastases (in the liver area) were detected in 6 patients; the remaining patients had no metastases. The degree of lymph node involvement in most patients was not determined, the ten corresponding N1. The control group consisted of 16 healthy people (50<!--> <!-->+<!--> <!-->6<!--> <!-->years old). Electric and viscoelastic Er parameters were investigated by dielectrophoresis, their membrane structure – by TLC and gas chromatography. The optical properties of BS were studied by the methods of ellipsometry. The reaction of the monoclonal antibody CD 24 with BS antigens of CC patients was studied by spectroscopic ellipsometry close to the conditions of surface plasmon resonance (SPR) (ProteOn XPR36 (BioRad).</p></div><div><h3>Results</h3><p>We observed significant differences in Er parameters, associated with the CC stage. Given in the 2nd group (T3–4) summarized rigidity, viscosity, electrical conductivity, the relative polarizability, indexes of aggregation and destruction were significantly higher than those in the 1st (T1–2) and in the control group (<em>p</em> <!--><<!--> <!-->0.001–0.05). At the same time the patients of the 2nd group had marked disturbances of Er deformability, leading to the development of microcirculatory disorders and tissue hypoxia with the expressed deficit of intracellular macroergs. We observed high levels of cholesterol fraction, oleic, stearic acids, high index of cholesterol/phospholipids (PHL) and low levels of total lipids, easily oxidable PHL, arachidonic acid, omega-3 index in Er membranes in the 2nd group in comparison with those in the 1st group of patients (<em>p</em> <!--><<!--> <!-->0.0001–0.03). Scanning ellipsometry showed marked heterogeneity in thickness and composition, the abundance of discontinuities in thin films of BS of patients in the 2nd group compared to the 1st one (<em>p</em> <!--><<!--> <!-->0.001). Increasing the refractive index in combination with the reduction in film thickness as CC stage was weighting has been observed (<em>p</em> <!--><<!--> <!-->0.01–0,<span><math><mrow><mo>⧹</mo></mrow></math></span>.05). The concentration of the antigens to the CD24 in the BS of patients (obtained by SPR) in the terminal stages of CC was higher than that in the T1–2 (<em>p</em> <!--><<!--> <!-->0.001). We revealed correlations between Er parameters, BS ellipsometry characteristics and biochemical parameters, which reflected the interaction between these components depending on the CC stage.</p></div><div>","PeriodicalId":11675,"journal":{"name":"Ejc Supplements","volume":"13 1","pages":"Pages 29-30"},"PeriodicalIF":0.0,"publicationDate":"2015-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.ejcsup.2015.08.052","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54309547","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}
Pub Date : 2015-11-01Epub Date: 2015-11-23DOI: 10.1016/j.ejcsup.2015.08.050
A. Kozlov
The hypothesis of the possible evolutionary role of tumors suggests that hereditary tumors may supply evolving multicellular organisms with extra cell masses for the expression of newly evolving genes (Kozlov, 2014). After expression of novel genes in tumor cells, tumors may differentiate in new directions and give rise to new cell types, tissues and organs.
In the presentation, the bulk of data supporting the positive evolutionary role of tumors will be reviewed, obtained both in the lab of the author and from the literature sources.
The following issues will be addressed: the widespread occurrence of tumors in multicellular organisms; features of tumors that could be used in evolution; the relationship of tumors to evo-devo; examples of recapitulation of some tumor features in recently evolved organs; the types of tumors that might play the role in evolution; examples of tumors that already have played the role in evolution.
The discussion of experimental confirmation of nontrivial predictions of the hypothesis will include the analysis of evolutionary novelty of tumor-specifically expressed EST sequences; ELFNI – AS1, a human gene with possible microRNA function expressed predominantly in tumors and originated in primates; PBOV1, a human gene of the recent de novo origin with predicted highly tumor-specific expression profile; and the evolutionary novelty of human cancer/testis antigen genes; the data obtained on transgenic fish tumors regression model; and other data.
It can be concluded that expression of protogenes, evolutionarily young and/or novel genes in tumors might be a new biological phenomenon, a phenomenon of carcino-evo-devo genes, predicted by the hypothesis of evolution by tumor neofunctionalization.
{"title":"A118","authors":"A. Kozlov","doi":"10.1016/j.ejcsup.2015.08.050","DOIUrl":"10.1016/j.ejcsup.2015.08.050","url":null,"abstract":"<div><p>The hypothesis of the possible evolutionary role of tumors suggests that hereditary tumors may supply evolving multicellular organisms with extra cell masses for the expression of newly evolving genes (Kozlov, 2014). After expression of novel genes in tumor cells, tumors may differentiate in new directions and give rise to new cell types, tissues and organs.</p><p>In the presentation, the bulk of data supporting the positive evolutionary role of tumors will be reviewed, obtained both in the lab of the author and from the literature sources.</p><p>The following issues will be addressed: the widespread occurrence of tumors in multicellular organisms; features of tumors that could be used in evolution; the relationship of tumors to evo-devo; examples of recapitulation of some tumor features in recently evolved organs; the types of tumors that might play the role in evolution; examples of tumors that already have played the role in evolution.</p><p>The discussion of experimental confirmation of nontrivial predictions of the hypothesis will include the analysis of evolutionary novelty of tumor-specifically expressed EST sequences; ELFNI – AS1, a human gene with possible microRNA function expressed predominantly in tumors and originated in primates; PBOV1, a human gene of the recent de novo origin with predicted highly tumor-specific expression profile; and the evolutionary novelty of human cancer/testis antigen genes; the data obtained on transgenic fish tumors regression model; and other data.</p><p>It can be concluded that expression of protogenes, evolutionarily young and/or novel genes in tumors might be a new biological phenomenon, a phenomenon of carcino-evo-devo genes, predicted by the hypothesis of evolution by tumor neofunctionalization.</p></div>","PeriodicalId":11675,"journal":{"name":"Ejc Supplements","volume":"13 1","pages":"Pages 28-29"},"PeriodicalIF":0.0,"publicationDate":"2015-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.ejcsup.2015.08.050","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54309521","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}
Pub Date : 2015-11-01Epub Date: 2015-11-23DOI: 10.1016/j.ejcsup.2015.08.049
S. Kovalenko , G. Paul , N. Matyash , A. Kozyakov
<div><p>Mutations in BRCA1 and CHEK2 genes associated with hereditary breast cancer were tested in 7920 randomly selected individuals of Novosibirsk (Russia). Mutations BRCA1 5382insC and CHEK2 1100delC were the most frequent, they were found in 0.25% and 0.4% of the general population respectively. We suggested to find mutations carriers by the screening of all breast/ovary cancer patients for the most frequent mutations (BRCA1 5382insC and CHEK2 1100delC) with subsequent analysis of the first-line relatives of cancer patients if one of the mutations was found.</p><p>From June 2013 till January 2015, all patients from Novosibirsk regional oncology hospital with the diagnosis of breast cancer and some patients with the diagnosis of ovary cancer were tested for mutations BRCA1 5382insC and CHEK2 1100delC. A total of 2655 cancer patients were analyzed independently of their family history. We found 122 mutations carriers, among them 99 patients with mutations in BRCA1 gene and 23 patients with mutation CHEK2 1100delC. Among mutation carriers, 105 patients agreed to have a medical genetic counseling and after pedigree analysis 193 first-line relatives aged above 25 years were elucidated. One hundred ten first-line relatives of mutation carriers were analyzed for the mutations presence and 40 mutations carriers were found among relatives.</p><p>From September 2013 till December 2013, 32 relatives of BRCA mutation carriers underwent breast MRI. In 5 cases, breast cancer was detected by MRI and all cancers except one were confirmed histologically with biopsy analysis. Importantly, all tumors were 5<!--> <!-->mm and less in size, stage I cancer was detected in all cases.</p><p>At a follow-up of 1.5<!--> <!-->years, all 105 mutation carrier probands were interviewed by phone regarding possible relapse and/or possible primary cancer in their relatives. Five of 105 probands lost to follow-up may have died. Among responding 100 patients, 2 died as reported by relatives, relapse was reported in 7 probands – mutation carrier probands, primary tumors were reported in 8 relatives of probands.</p><p>Mutation carrier probands reported one bilateral breast cancer, four ovary cancers, one bladder cancer and one non-specified oncogynecological tumor.</p><p>There were five cases of primary breast cancer, one ovary cancer, one colon cancer, one lung cancer among relatives of breast cancer patients with mutations. The frequency of tumors found in mutation carriers exceeded the average frequencies of cancer for this population.</p><p>The economic value of the regional genetic screening can be easily estimated according to the data obtained in this study and data on treatment cost for stage I and stage IV breast cancer. To summarize briefly, the screening of hot-spot mutations provides not only increase of lifespan expectancy and life quality for mutation carriers, but can be also a tool for financial saving of medical system due to the increase of early stage breast can
{"title":"P107","authors":"S. Kovalenko , G. Paul , N. Matyash , A. Kozyakov","doi":"10.1016/j.ejcsup.2015.08.049","DOIUrl":"10.1016/j.ejcsup.2015.08.049","url":null,"abstract":"<div><p>Mutations in BRCA1 and CHEK2 genes associated with hereditary breast cancer were tested in 7920 randomly selected individuals of Novosibirsk (Russia). Mutations BRCA1 5382insC and CHEK2 1100delC were the most frequent, they were found in 0.25% and 0.4% of the general population respectively. We suggested to find mutations carriers by the screening of all breast/ovary cancer patients for the most frequent mutations (BRCA1 5382insC and CHEK2 1100delC) with subsequent analysis of the first-line relatives of cancer patients if one of the mutations was found.</p><p>From June 2013 till January 2015, all patients from Novosibirsk regional oncology hospital with the diagnosis of breast cancer and some patients with the diagnosis of ovary cancer were tested for mutations BRCA1 5382insC and CHEK2 1100delC. A total of 2655 cancer patients were analyzed independently of their family history. We found 122 mutations carriers, among them 99 patients with mutations in BRCA1 gene and 23 patients with mutation CHEK2 1100delC. Among mutation carriers, 105 patients agreed to have a medical genetic counseling and after pedigree analysis 193 first-line relatives aged above 25 years were elucidated. One hundred ten first-line relatives of mutation carriers were analyzed for the mutations presence and 40 mutations carriers were found among relatives.</p><p>From September 2013 till December 2013, 32 relatives of BRCA mutation carriers underwent breast MRI. In 5 cases, breast cancer was detected by MRI and all cancers except one were confirmed histologically with biopsy analysis. Importantly, all tumors were 5<!--> <!-->mm and less in size, stage I cancer was detected in all cases.</p><p>At a follow-up of 1.5<!--> <!-->years, all 105 mutation carrier probands were interviewed by phone regarding possible relapse and/or possible primary cancer in their relatives. Five of 105 probands lost to follow-up may have died. Among responding 100 patients, 2 died as reported by relatives, relapse was reported in 7 probands – mutation carrier probands, primary tumors were reported in 8 relatives of probands.</p><p>Mutation carrier probands reported one bilateral breast cancer, four ovary cancers, one bladder cancer and one non-specified oncogynecological tumor.</p><p>There were five cases of primary breast cancer, one ovary cancer, one colon cancer, one lung cancer among relatives of breast cancer patients with mutations. The frequency of tumors found in mutation carriers exceeded the average frequencies of cancer for this population.</p><p>The economic value of the regional genetic screening can be easily estimated according to the data obtained in this study and data on treatment cost for stage I and stage IV breast cancer. To summarize briefly, the screening of hot-spot mutations provides not only increase of lifespan expectancy and life quality for mutation carriers, but can be also a tool for financial saving of medical system due to the increase of early stage breast can","PeriodicalId":11675,"journal":{"name":"Ejc Supplements","volume":"13 1","pages":"Page 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.2015.08.049","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54309506","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}
Pub Date : 2015-11-01Epub Date: 2015-11-23DOI: 10.1016/j.ejcsup.2015.08.030
E. Gashenko , V. Lebedeva , E. Tsykalenko , G. Russkikh , I. Brak , T. Korolenko
<div><h3>Background</h3><p>The research for procathepsin B and endogenous inhibitors of cysteine proteases in tumor markers of human reproductive system is important for early diagnostics of cancer. Preform of cathepsin B and cystatins B and C also are universally involved into development of different tumors. Tumor cells as well as tumor-associated macrophages have been shown to secrete active forms of proteases and their inhibitors; however, their roles, especially those of proenzymes as markers of malignancy, are still under investigation.</p></div><div><h3>Aim</h3><p>to evaluate procathepsin B and endogenous inhibitors of cysteine proteases cystatins B and C in tumors of reproductive system.</p></div><div><h3>Materials and methods</h3><p>Serum and ascites fluid of 38 patients with ovarian cancer (among them 15 patients after treatment) and benign ovarian tumors (<em>n</em> <!-->=<!--> <!-->9), endometrial cancer (<em>n</em> <!-->=<!--> <!-->31, before treatment 14), mammalian cancer (<em>n</em> <!-->=<!--> <!-->29, before treatment 18) of stages II–IV for all groups, from Department of Gynecology of Regional Oncology Center, Novosibirsk, were under investigation. Serum of practically healthy women aged 18–80 (<em>n</em> <!-->=<!--> <!-->82) from Regional Diagnostic Center, Novosibirsk, was used as a control group. Serum of women with tumors of the reproductive system and ascites fluids of women with ovarian tumors (aged 18–80 years), before operation were used for assay of procathepsin B, cysteine protease inhibitors cystatins B and C. Serum procathepsin B concentration was measured by ELISA commercial kit for human (R&D) USA; cystatin C using BioVendor commercial kits (Czechia), cystatin B – with help of ELISA kits for human (USCN Life Science Inc., Wuhan, China). Common biomarker of ovarian cancer, CA-125, was assayed by using a commercial kit (Vector, Koltsovo, Novosibirsk Region, Russia). Statistical analysis performed by one a way ANOVA Statistic 12, program with help of Kruskall–Wallis test and the Mann–Whitney <em>U</em> test used to assess differences in procathepsin B or cystatins B and C levels between patient groups. Statistical analysis was performed with the soft package Statistics 12, with the level of significance being set at <0.05.</p></div><div><h3>Results</h3><p>In serum of patients with endometrial cancer, ovarian cancer, mammalian cancer – significant increases in serum procathepsin B (<em>p</em> <!--><<!--> <!-->0.001), cystatin B (<em>p</em> <!--><<!--> <!-->0.05) and CA-125 (<em>p</em> <!--><<!--> <!-->0.001) were noted. However, in patients with benign tumor increased serum common tumor marker CA-125 (<em>p</em> <!-->=<!--> <!-->0.005 vs. healthy controls) was shown without any changes in serum level of procathepsin B, cystatins B and C. Concentrations procathepsin B and Cystatin B in serum and ascites of patients with ovarian tumor was used to assess differences in procathepsin B or cystatins B and C
{"title":"A91","authors":"E. Gashenko , V. Lebedeva , E. Tsykalenko , G. Russkikh , I. Brak , T. Korolenko","doi":"10.1016/j.ejcsup.2015.08.030","DOIUrl":"10.1016/j.ejcsup.2015.08.030","url":null,"abstract":"<div><h3>Background</h3><p>The research for procathepsin B and endogenous inhibitors of cysteine proteases in tumor markers of human reproductive system is important for early diagnostics of cancer. Preform of cathepsin B and cystatins B and C also are universally involved into development of different tumors. Tumor cells as well as tumor-associated macrophages have been shown to secrete active forms of proteases and their inhibitors; however, their roles, especially those of proenzymes as markers of malignancy, are still under investigation.</p></div><div><h3>Aim</h3><p>to evaluate procathepsin B and endogenous inhibitors of cysteine proteases cystatins B and C in tumors of reproductive system.</p></div><div><h3>Materials and methods</h3><p>Serum and ascites fluid of 38 patients with ovarian cancer (among them 15 patients after treatment) and benign ovarian tumors (<em>n</em> <!-->=<!--> <!-->9), endometrial cancer (<em>n</em> <!-->=<!--> <!-->31, before treatment 14), mammalian cancer (<em>n</em> <!-->=<!--> <!-->29, before treatment 18) of stages II–IV for all groups, from Department of Gynecology of Regional Oncology Center, Novosibirsk, were under investigation. Serum of practically healthy women aged 18–80 (<em>n</em> <!-->=<!--> <!-->82) from Regional Diagnostic Center, Novosibirsk, was used as a control group. Serum of women with tumors of the reproductive system and ascites fluids of women with ovarian tumors (aged 18–80 years), before operation were used for assay of procathepsin B, cysteine protease inhibitors cystatins B and C. Serum procathepsin B concentration was measured by ELISA commercial kit for human (R&D) USA; cystatin C using BioVendor commercial kits (Czechia), cystatin B – with help of ELISA kits for human (USCN Life Science Inc., Wuhan, China). Common biomarker of ovarian cancer, CA-125, was assayed by using a commercial kit (Vector, Koltsovo, Novosibirsk Region, Russia). Statistical analysis performed by one a way ANOVA Statistic 12, program with help of Kruskall–Wallis test and the Mann–Whitney <em>U</em> test used to assess differences in procathepsin B or cystatins B and C levels between patient groups. Statistical analysis was performed with the soft package Statistics 12, with the level of significance being set at <0.05.</p></div><div><h3>Results</h3><p>In serum of patients with endometrial cancer, ovarian cancer, mammalian cancer – significant increases in serum procathepsin B (<em>p</em> <!--><<!--> <!-->0.001), cystatin B (<em>p</em> <!--><<!--> <!-->0.05) and CA-125 (<em>p</em> <!--><<!--> <!-->0.001) were noted. However, in patients with benign tumor increased serum common tumor marker CA-125 (<em>p</em> <!-->=<!--> <!-->0.005 vs. healthy controls) was shown without any changes in serum level of procathepsin B, cystatins B and C. Concentrations procathepsin B and Cystatin B in serum and ascites of patients with ovarian tumor was used to assess differences in procathepsin B or cystatins B and C ","PeriodicalId":11675,"journal":{"name":"Ejc Supplements","volume":"13 1","pages":"Pages 16-17"},"PeriodicalIF":0.0,"publicationDate":"2015-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.ejcsup.2015.08.030","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54309061","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}
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