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研究生:林巧威
研究生(外文):Chiao-Wei Lin
論文名稱:Stat5訊息傳導與SMN2基因全長型式轉錄產物的關係研究
論文名稱(外文):Involvement of Stat5 signaling pathway in SMN2 exon 7 inclusion
指導教授:李鴻李鴻引用關係沈哲鯤沈哲鯤引用關係
指導教授(外文):Hung-LiChe-Kun James Shen
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:遺傳學研究所
學門:生命科學學門
學類:生物訊息學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
中文關鍵詞:脊髓肌肉萎縮症
外文關鍵詞:spinal muscular atrophySMNSMAStat5
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脊髓肌肉萎縮症(SMA)是一種常見的隱性體染色體遺傳疾病,特徵為脊髓的運動神經元死亡造成四肢及軀幹的麻痹而導致肌肉萎縮。目前已知survival of motor neuron 1 (SMN1)基因是SMA的決定基因。在人類的基因體中, SMN1和SMN2有95%的相似性,能產生相同的蛋白質,但大部份的患者都是SMN1基因有homozygous突變或缺失。SMN1經過轉錄後的產物幾乎都是全長mRNA,並可經轉譯做出有功能的蛋白質;而SMN2只能轉錄出少量的全長mRNA,主要都是缺少exon7(∆exon7)的mRNA,∆exon7 mRNA經轉錄而做出C端截短而容易被分解的蛋白質。之前,我們將人類的SMN2的genomic DNA轉殖到老鼠的似運動神經元的細胞株(NSC34),並且利用此細胞株做了一系列的藥物篩選。我們篩選出三種可有效改變SMN2的剪接作用和增加SMN2表現的藥物,分別是sodium vanadate,aclarubicin以及trichostatin A。本研究發現,當這三種藥物在分別處理SMN2-transfected NSC34後,細胞的Stat5 signaling會被活化。當我們在SMN2-transfected NSC34表現constitutively active Stat5 mutant (STAT5A1*6)後,可增加SMN2轉錄出全長mRNA的能力。另外,我們利用gems的數目做為有功能的SMN蛋白質數量的指標。在SMA病人的淋巴細胞表現STAT5A1*6可增加gems的數目。我們認為Stat5 signaling參與了這三種藥物增加SMN2表現的機制。此外,活化Stat5 signaling可增強SMN2轉錄的能力,並且增加有功能的SMN蛋白質的量。活化Stat5 signaling或許可利用來做為治療SMA的一種途徑。
Spinal muscular atrophy (SMA) is a recessive disease characterized by degeneration of motor neuron in the spinal cord. SMA is caused by homologous mutation of the survival of motor neuron 1 (SMN1) gene, which exists in another nearly identical copy, SMN2. Although SMN1 and SMN2 potentially encode identical proteins, main transcripts of SMN1 are functional full-length mRNA, whereas the predominant transcripts produced by SMN2 are ∆exon7. We transfected the SMN2 genomic DNA into NSC34, a motor neuron-like mouse cell line, to establish a drug screening system for SMN2 expression pattern. In our previous study, sodium vanadate, aclarubicin and trichostatin A changed the expression ratio of full-length to ∆exon7 mRNA of SMN2 in SMN2-transfected NSC34. In this report, I demonstrated that activation of Stat5 signaling is the common event by treated with the drugs, respectively. Expression of the constitutively active Stat5 mutant, STAT5A1*6, raised the level of full-length SMN mRNA. Besides, expression of STAT5A1*6 increased the number of nuclear gems which are functional indicators of SMN in lymphocytes from SMA patients. The results revealed that activation of Stat5 signaling raised full-length mRNA and functional SMN of SMN2. I propose that Stat5 signaling is involved in changing the ratio of full-length to ∆exon7 mRNA of SMN2. Stat5 signaling may play a role for SMA treatment.
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