胶质母细胞瘤中离子通道的预后价值及KCNB1影响自噬的研究

发布时间：2018-01-29 07:11

  本文关键词： 原发性胶质母细胞瘤 RNA测序数据 危险评分 离子通道胶质瘤 KCNB1 离子通道 自噬　出处：《南方医科大学》2017年博士论文　论文类型：学位论文

【摘要】：第一部分基于RNA测序数据的3个离子通道基因组成的危险评分在原发性胶质母细胞瘤的预后价值背景:近年来研究表明,离子通道基因在胶质瘤的进展中起着重要作用。本研究目的在于应用RNA测序数据筛选出能够预测原发性胶质母细胞瘤(pGBM)预后的基于离子通道基因的危险评分。方法:通过单变量COX及t检验在中国脑胶质瘤大样本库(CGGA)各个级别的胶质瘤中鉴定出3个与预后和级别相关的离子通道基因,并基于这3个基因构建危险评分,赋予每个pGBM患者一个评分。根据评分中位值,将83例pGBM患者分成高危险评分组(n=42)和低危险评分组(n=41)。并利用基因功能分析(GO,Gene ontology)及基因富集分析(GSEA,Gene Set Enrichment Analysis)进一步研究危险评分相关的生物学功能。这些结果均在另外两个数据库中进行验证(TCGA和REMBRANDT)。结果:我们筛选出3个与胶质瘤预后及级别相关的离子通道基因(KCNN4、KCNB1和KCNJ10),并且构建了基于这3个基因表达值的危险评分。生存分析发现相比较低危险评分组,高危险评分组患者预后要差,对化疗反应敏感,且COX多元回归分析证实此危险评分是一个独立的预后预测因子。亚型分析显示高危险评分组倾向于胶质瘤Mesenchymal型和IDH1野生型。GO分析显示高危险评分组倾向于凋亡和血管生成等功能。GSEA分析发现高危险评分组倾向于细胞增殖和运动行为等功能。以上结果均在另外两个数据库中得到验证(TCGA 和 REMBRANDT)。结论:这个危险评分系统能更好的预测pGBM患者的预后,并有助于胶质瘤患者的个体化治疗。第二部分电压门控性钾离子通道KCNB1对胶质瘤预后的影响及自噬机制研究背景:第一部分研究发现KCNB1与胶质瘤级别及预后相关,本课题进一步研究KCNB1影响胶质瘤进展的分子机制。方法:本研究收集42例胶质瘤独立标本进行qRT-PCR验证,并收集41例Ⅲ级,83例Ⅳ级胶质瘤标本进行Kaplan-Meier生存分析。整合系统生物学预测KCNB1的功能,然后采用免疫印迹、免疫荧光、流式细胞术、MTT等技术,通过体内外实验去验证。结果:qRT-PCR验证KCNB1表达与级别呈负相关,且生存分析表明KCNB1高表达组有较好的总生存期及无瘤生存期。生物学分析及实验证实KCNB1具有促进自噬、凋亡,抑制增殖、侵袭功能。KCNB1通过激活ERK激酶促进自噬,进一步用U0126和siERK1/2去阻滞ERK会使自噬减少。小鼠成瘤实验也证明了 KCNB1抑制增殖和促进自噬的功能。结论:KCNB1基因激活ERK通路促进自噬,从而抑制胶质瘤的生长。
[Abstract]:Part I the prognostic value of risk scores for the composition of three ion channel genes based on RNA sequencing data in primary glioblastoma: recent studies have shown. Ion channel genes play an important role in the progression of gliomas. The aim of this study was to screen out pGBM, which can predict primary glioblastoma by using RNA sequencing data. Prognostic risk scores based on ion channel genes. Methods: single variable COX and t test were performed in a large sample library of gliomas in China. Three ion channel genes associated with prognosis and grade were identified in each grade of glioma. Risk scores were constructed based on these three genes, and each pGBM patient was given a score, according to the median value of the score. 83 patients with pGBM were divided into high risk score group (n = 42) and low risk score group (n = 41). Gene ontologyand gene enrichment analysis. Gene Set Enrichment analysis further studies the biological functions associated with hazard scores. These results are validated in two other databases. Results: we screened three ion channel genes (KCNN4) associated with prognosis and grade of glioma. KCNB1 and KCNJ10, and constructed a risk score based on these three gene expression values. Survival analysis found that compared with the low risk score group, the prognosis of patients with high risk score group was poor. Sensitive to chemotherapy. COX multiple regression analysis confirmed that the risk score was an independent prognostic predictor. Subtype analysis showed that the high risk score group tended to glioma Mesenchymal type and IDH1 wild type. G. O analysis showed that the high risk score group tended to function such as apoptosis and angiogenesis. GSEA analysis showed that the high risk score group tended to function such as cell proliferation and motor behavior. All the above results were obtained in the other two databases. To validation. TCGA and REMBRAN DT.Conclusion: this risk scoring system can better predict the prognosis of patients with pGBM. The second part is the effect of voltage-gated potassium channel (KCNB1) on the prognosis of glioma and the mechanism of autophagy. In the first part, KCNB1 was found to be associated with glioma grade and prognosis. In this study, we further studied the molecular mechanism of KCNB1 affecting glioma progression. Methods: 42 cases of glioma were collected for qRT-PCR verification and 41 cases for grade 鈪，

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