使用MicroRNA微陣列分析來觀察在惡性膠原母細胞瘤中整體MicroRNAs的表現情況

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题名:	使用MicroRNA微陣列分析來觀察在惡性膠原母細胞瘤中整體MicroRNAs的表現情況 Expression Profile of MicroRNAs in Glioblastoma Multiforme Tissues
作者:	李宗祐
贡献者:	免疫學研究所碩士班
关键词:	惡性膠原母細胞瘤;微小RNA GBM;microRNA
日期:	2011-07-25
上传时间:	2011-10-17 16:36:05 (UTC+8)
出版者:	中國醫藥大學
摘要:	惡性膠原母細胞瘤(GBM)是一種具高度侵襲性的常見腦部腫瘤，即使結合許多有效的抗癌治療，平均存活時間仍然只有大約一年。MicroRNAs是一種細胞原生的小片段RNA，且不帶有轉譯編碼，並藉由結合mRNA 3’UTR區域的方式來減少目標基因的蛋白質表現。近來的研究發現microRNAs在惡性膠原母細胞瘤的許多生物性功能上中扮演了相當重要的調節角色，包含了癌細胞增生、癌細胞轉移、癌幹細胞生長及血管新生。在這個研究中，希望藉由比較microRNAs在正常腦組織和惡性膠原母細胞瘤之間表現情況的差異來找出表現量在兩者間有異常變化的microRNAs。利用microRNAs微陣列分析同時監測716個microRNAs在七個臨床病人檢體、兩個臨床檢體的初帶培養細胞及一個正常腦組織的表現情形。從結果中得知，有十三個microRNAs在惡性膠原母細胞瘤檢體中具有顯著的表現量下降。利用即時定量聚合酶連鎖反應(Real-Time PCR)對這十三個microRNAs的微陣列分析的結果確認，結果只有部分microRNAs的表現情況較吻合，其中miR-137和miR-7在各檢體中皆完全符合。而這些microRNAs具有成為在惡性膠原母細胞瘤上的biomark的潛力。為了進一步研究這些microRNAs所影響到的生物性功能，結合microRNAs目標基因的預測網站及惡性膠原母細胞瘤基因表現的資料庫，預測出了數個microRNAs的目標基因，在Real-Time PCR的結果中以TBX2基因為miR-7的目標基因的可能性最大，未來將更加的證實TBX2與miR-7的關係，並找出TBX2與miR-7在惡性膠原母細胞瘤中所扮演的角色。 Glioblastoma multiforme (GBM), is highly aggressive with a median survival of one year, even with the most effective combined therapies. MicroRNAs (miRNAs) are short endogenous noncoding RNAs that silence the production of proteins by binding to mRNA 3’UTR regions. miRNAs have been shown their roles in the regulation of biological properties in glioblastomas, including cell proliferation, invasion, cancer stem cell growth, and angiogenesis. In this study, we compared the expression levels of microRNAs between clinic GBM tissues and normal brain tissues. For acquiring the profiles of different expression levels of global miRNAs, 7 clinic samples from GBM patients, 1 normal brain tissue and 2 primary cells derived from GBM tissues were processed and analyzed by use of the miRNAs microarray system. In our results, expression levels of 13 microRNAs were shown to downregulate significantly in GBM tissue samples. These results from miRNA array analysis were also confirmed by use of qRT-PCR analysis. The expression levels of 6 miRNAs were the same as the findings in the miRNA array system. In summary, a panel of differentially expressed miRNAs derived from the microRNAs expression profile may serve as the potential molecular biomarkers for the therapy target of glioblastoma. By use of computational algorithm, we further predicted potent genes which are targeted by 13 miRNAs. And use published DNA microarray database in GBM to distinguish the genes affected by miRNAs. Some of predicted genes had been shown to play key functions in proliferation, cell cycle, development, cell survival and migration. And further studies to investigate the effect of these miRNAs on glioblastomas biology are necessary.
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