本文關(guān)鍵詞： 原發(fā)性膠質(zhì)母細(xì)胞瘤 RNA測序數(shù)據(jù) 危險(xiǎn)評分 離子通道膠質(zhì)瘤 KCNB1 離子通道 自噬　出處：《南方醫(yī)科大學(xué)》2017年博士論文　論文類型：學(xué)位論文

【摘要】：第一部分基于RNA測序數(shù)據(jù)的3個(gè)離子通道基因組成的危險(xiǎn)評分在原發(fā)性膠質(zhì)母細(xì)胞瘤的預(yù)后價(jià)值背景:近年來研究表明,離子通道基因在膠質(zhì)瘤的進(jìn)展中起著重要作用。本研究目的在于應(yīng)用RNA測序數(shù)據(jù)篩選出能夠預(yù)測原發(fā)性膠質(zhì)母細(xì)胞瘤(pGBM)預(yù)后的基于離子通道基因的危險(xiǎn)評分。方法:通過單變量COX及t檢驗(yàn)在中國腦膠質(zhì)瘤大樣本庫(CGGA)各個(gè)級別的膠質(zhì)瘤中鑒定出3個(gè)與預(yù)后和級別相關(guān)的離子通道基因,并基于這3個(gè)基因構(gòu)建危險(xiǎn)評分,賦予每個(gè)pGBM患者一個(gè)評分。根據(jù)評分中位值,將83例pGBM患者分成高危險(xiǎn)評分組(n=42)和低危險(xiǎn)評分組(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)進(jìn)一步研究危險(xiǎn)評分相關(guān)的生物學(xué)功能。這些結(jié)果均在另外兩個(gè)數(shù)據(jù)庫中進(jìn)行驗(yàn)證(TCGA和REMBRANDT)。結(jié)果:我們篩選出3個(gè)與膠質(zhì)瘤預(yù)后及級別相關(guān)的離子通道基因(KCNN4、KCNB1和KCNJ10),并且構(gòu)建了基于這3個(gè)基因表達(dá)值的危險(xiǎn)評分。生存分析發(fā)現(xiàn)相比較低危險(xiǎn)評分組,高危險(xiǎn)評分組患者預(yù)后要差,對化療反應(yīng)敏感,且COX多元回歸分析證實(shí)此危險(xiǎn)評分是一個(gè)獨(dú)立的預(yù)后預(yù)測因子。亞型分析顯示高危險(xiǎn)評分組傾向于膠質(zhì)瘤Mesenchymal型和IDH1野生型。GO分析顯示高危險(xiǎn)評分組傾向于凋亡和血管生成等功能。GSEA分析發(fā)現(xiàn)高危險(xiǎn)評分組傾向于細(xì)胞增殖和運(yùn)動(dòng)行為等功能。以上結(jié)果均在另外兩個(gè)數(shù)據(jù)庫中得到驗(yàn)證(TCGA 和 REMBRANDT)。結(jié)論:這個(gè)危險(xiǎn)評分系統(tǒng)能更好的預(yù)測pGBM患者的預(yù)后,并有助于膠質(zhì)瘤患者的個(gè)體化治療。第二部分電壓門控性鉀離子通道KCNB1對膠質(zhì)瘤預(yù)后的影響及自噬機(jī)制研究背景:第一部分研究發(fā)現(xiàn)KCNB1與膠質(zhì)瘤級別及預(yù)后相關(guān),本課題進(jìn)一步研究KCNB1影響膠質(zhì)瘤進(jìn)展的分子機(jī)制。方法:本研究收集42例膠質(zhì)瘤獨(dú)立標(biāo)本進(jìn)行qRT-PCR驗(yàn)證,并收集41例Ⅲ級,83例Ⅳ級膠質(zhì)瘤標(biāo)本進(jìn)行Kaplan-Meier生存分析。整合系統(tǒng)生物學(xué)預(yù)測KCNB1的功能,然后采用免疫印跡、免疫熒光、流式細(xì)胞術(shù)、MTT等技術(shù),通過體內(nèi)外實(shí)驗(yàn)去驗(yàn)證。結(jié)果:qRT-PCR驗(yàn)證KCNB1表達(dá)與級別呈負(fù)相關(guān),且生存分析表明KCNB1高表達(dá)組有較好的總生存期及無瘤生存期。生物學(xué)分析及實(shí)驗(yàn)證實(shí)KCNB1具有促進(jìn)自噬、凋亡,抑制增殖、侵襲功能。KCNB1通過激活ERK激酶促進(jìn)自噬,進(jìn)一步用U0126和siERK1/2去阻滯ERK會(huì)使自噬減少。小鼠成瘤實(shí)驗(yàn)也證明了 KCNB1抑制增殖和促進(jìn)自噬的功能。結(jié)論:KCNB1基因激活ERK通路促進(jìn)自噬,從而抑制膠質(zhì)瘤的生長。
[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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