E. Rybalkina , G. Pavlova , N. Moiseeva , O. Susova , A. Mitrofanov , D. Panteleev , N. Pustogarov
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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":0.0000,"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":"0","resultStr":"{\"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\":null,\"url\":null,\"abstract\":\"<div><h3>Background</h3><p>Glioblastomas (GBL) are most aggressive brain tumors in adults. They are often radio- and chemotherapy resistant. 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引用次数: 0
摘要
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。
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).
期刊介绍：
EJC Supplements is an open access companion journal to the European Journal of Cancer. As an open access journal, all published articles are subject to an Article Publication Fee. Immediately upon publication, all articles in EJC Supplements are made openly available through the journal''s websites.
EJC Supplements will consider for publication the proceedings of scientific symposia, commissioned thematic issues, and collections of invited articles on preclinical and basic cancer research, translational oncology, clinical oncology and cancer epidemiology and prevention.
Authors considering the publication of a supplement in EJC Supplements are requested to contact the Editorial Office of the EJC to discuss their proposal with the Editor-in-Chief.
EJC Supplements is an official journal of the European Organisation for Research and Treatment of Cancer (EORTC), the European CanCer Organisation (ECCO) and the European Society of Mastology (EUSOMA).
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