Micro-RNAs在惡性神經膠質母細胞瘤與正常腦細胞中之不同表現

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Title:	Micro-RNAs在惡性神經膠質母細胞瘤與正常腦細胞中之不同表現 Micro-RNAs Express Differentially in Glioblastoma Multiforme and Normal Brain Cells
Authors:	莊皓宇;Hao-Yu Chuang
Contributors:	醫學院臨床醫學研究所碩士班
Keywords:	神經腫瘤幹細胞;惡性神經膠質瘤;免疫細胞治療;Astrocytoma;Gene expression assay;GBM;Glioblastoma multiforme;Micro-RNA;Micro-array;micro-RNA chip
Date:	2009
Issue Date:	2010-09-29 12:17:14 (UTC+8)
Abstract:	試驗主題本研究主要是研究惡性神經膠質瘤中，microRNA於神經腫瘤細胞及神經腫瘤幹細胞中扮演之角色。我們以組織庫挑選收集病人檢體(已獲得病人同意後)，包含惡性神經膠質瘤和正常腦組織檢體細胞，篩檢出有意義的micro-RNA，利用GBM tissue microRNA Chip Data Analysis 瞭解惡性神經膠質瘤和正常腦組織細胞中microRNA之表現及差異程度。研究背景及目的即使最先進之治療改變包括先進手術導航手術方法，化學治療，放射線治療，對於惡性神經膠質瘤之預後，進展仍十分有限。只能多延長3至6個月生命，腫瘤幹細胞雖然只佔1-2%之惡性腫瘤，它卻有堅強生長分化能力，且對臨床之治療有頑強之抗性。它具有強大自我更新及DNA修護能力，我們認為microRNA在神經腫瘤細胞及神經腫瘤幹細胞中之角色扮演極為重要，有急需研究瞭解之必要。目前已知在不同狀態下的神經腫瘤細胞及神經腫瘤幹細胞(CD133+, CD133-)之中，會有不同的microRNA表現，如能知道microRNA影響神經腫瘤細胞生長分化因子，就可以擬定治療改進之對策，以增進惡性神經膠質瘤預後。實驗設計本實驗以組織庫挑選收集病人檢體((檢體6件，實驗檢體取自本院組織庫中已獲得病人同意之檢體，包含惡性神經膠質瘤和正常腦組織等四大類的檢體。) ，包含惡性神經膠質瘤和正常腦組織檢體，利用GBM tissue microRNA Chip Data Analysis 瞭解惡性神經膠質瘤和正常腦組織細胞中microRNA之表現及差異程度。我們統計並比較microRNA 在不同檢體中的表現差異(miRNA intensity<1, adjust to 1, Ratio=log2 (case#1/case#2) ), 此外我們也利用分析出來的資料, 利用 Gene expression assay datas of Normal Brain Tissue ( GDS596 ) 做基因庫的分析( from Affymetrix U133A and GBM primary culture cell lines (Affymetrix U133 Plus 2.0)). 我們發現有些人類細胞或病毒株之microRNA 具有相當意義之正調控或負調控的表現, 而這些具統計意義之表現存在於惡性神經膠質瘤和正常腦組織細胞之中. 結論我們可以初步證實microRNA在惡性腦瘤幹細胞中扮演的角色 , 並期待microRNA正調控或負調控的表現, 可以影響惡性神經膠質瘤細胞之表現，並期待可以發展可行之治療策略，以增進治療惡性神經膠質瘤之臨床治療預後。 Object We hypothesize that Micro-RNAs might play an important roles in glioblastoma glastoma (GBM) cells, and differentially express between GBM and normal brain cells. We compare clinical datas using MicroRNA Chip Data Analysis and statistical analyses by use of the algorithm in miRBase. Methods We compare clinical datas using MicroRNA Chip Data Analysis and statistical analyses by use of the algorithm in miRBase. After surgical resection, we make definite GBM pathological specimen examination and to culture GBM cells. The normal brain tissue was donated from a patient expired with intracranial lesions, and we excise the normal brain tissue within 30 minutes just after his death. We want to find some differently expressed miRs (If miRNA intensity<1, adjust to 1, Ratio=log2 (case#1/case#2) ) in these 4 types of cells, and adopt to molecular- biological research for malignant brain tumors. Furthermore, we also adopt our preliminary datas to compare with Gene expression assay datas of Normal Brain Tissue ( GDS596 ) from Affymetrix U133A and GBM primary culture cell lines (Affymetrix U133 Plus 2.0) Results We have obtained a promising preliminary miRNA profiling which resulted from comparing with GBM and non-tumor brain tissues. By use of the algorithm in miRBase Targets, some miRNAs have been computationally predicted to show the significantly change of expression ratio in our preliminary miRNA data. Conclusion Some Micro-RNAs significantly expressed differentially in GBM cells and normal brain cells. Some express as up-regulation, and others express as down-regulation. Further study including Micro-RNAs RT- PCR and gene expression assay are ongoing. Micro-RNAs might play an important
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