中間絲（IF）是細胞對抗機械應力一個主要架構，然而在核轉錄的調控的新功能，在癌症研究中越來越多的關注。這種調控已經被認為是幹細胞發育的關鍵步驟。我們最近的研究也證實了鱗片狀細胞口腔癌中角蛋白融合的存在與腫瘤大小，轉移和總生存相關。在所鑑定的融合體中，發現KRT6-KRT14融合體V 7（KRT6-KRT14/V7）在細胞核周圍形成與核變形和微核形成相關的過度活躍的顆粒狀聚集體。在Time-lapse及免疫染色研究進一步證實K6-K14 / V7可以干擾LINC complex (核骨架和細胞骨架的連接子）的結構，導致染色體分離不均勻並活化 DNA 損害反應造成細胞凋零死亡。有趣的是，隨著p53-p21自我監測中的遺傳缺陷，長期的基因打開效應可能引發基因改變的積累。未來此計畫還將討論涉及由KRT6-KRT14/V7介導的核機械轉導的幾個關鍵途徑，我們的研究結果揭示了在癌症發展中起作用的角蛋白融合的獨特功能。

Intermediate filaments (IFs), a major structural architecture in cells against mechanical stresses, gain more attention in cancer research due to thei novel functions in regulation of nuclear mechano-transduction. Such regulation has been noted as a key step for stem cell development and tissue homeostasis. Our recent studies also confirmed the existence of keratin fusions in OSCCs that correlate with tumor size, metastasis and overall survival. Among the fusions identified, the variant 7 of K6-K14 fusions (K6-K14/V7) was found to form over-active granule-like aggregates surrounding the cell nucleus that associate with nuclear deformation and micronucleus formation. Time-lapse study further confirmed that the K6-K14/V7 can disturb the structure of LINC complex (linker of nucleoskeleton and cytoskeleton), resulting in uneven chromosome segregation followed by cell apoptosis via activating DNA damage responses. Interestingly, genetic defects in p53-p21 self-surveillance. Long-term gene-turned on can trigger accumulations of genetic alterations. Several key pathways involved in nuclear mechano-transduction mediated by K6-K14/V7 will be also discussed. Our findings uncover a unique function of keratin fusions that plays roles in cancer development.

論文審定書-----------------------------------------------------------------------------------------ii
誌謝 (Ackowledgements) -----------------------------------------------------------------------iii
中文摘要 (Chinese of abstract) ---------------------------------------------------------------- iv
英文摘要 (English of abstract) -----------------------------------------------------------------v
縮寫表(Abbreviations) ---------------------------------------------------------------------------vi
前言 (Introduction) -------------------------------------------------------------------------------1
材料與方法 (Materials and Methods) ---------------------------------------------------------5
實驗結果 (Results)--------------------------------------------------------------------------------9
討論 (Discussion)--------------------------------------------------------------------------------13
圖表 (Figures)------------------------------------------------------------------------------------16
圖1、識別染色體易位t(12; 17)(q13;q21)在口腔鱗狀上皮細胞癌(OSCC) --------16
圖2、將基因融合KRT6-KRT14轉錄在口腔鱗狀細胞癌進行鑑定-----------------17
圖3、臨床資料中顯示KRT6-KRT14基因融合變異與口腔鱗狀細胞癌之相關----18
圖4、KRT6-KRT14基因融合變異與口腔鱗狀細胞癌細胞關係----------------------19
圖5、K6-K14基因融合V7型態基因建構-------------------------------------------------20
圖6、角蛋白融合V7型態及在細胞內核變形和微核形成-----------------------------21
圖7、角蛋白融合V7型態及在細胞內核變形和微核形成-----------------------------22
圖8、角蛋白融合V7表達誘導DNA鏈斷裂並和細胞生長抑制---------------------23
圖9、角蛋白融合V7激活發 DNA損傷反應--------------------------------------------24
圖10、角蛋白融合V7激活發 DNA損傷反應使用檢測其路徑指標(1)-------------25
圖11、角蛋白融合V7激活發 DNA損傷反應使用檢測其路徑指標(2)-------------26
圖12、ATM抑制劑及p53突變逆轉KRT6-KRT14/V7誘導的細胞生長停滯---27
圖13、K6-K14 / V7通過中心體改變干擾細胞分裂------------------------------------28
圖14、K6-K14 / V7通過打亂LINC複合體導致細胞核的不穩定--------------------29
圖15、K6-K14 / V7通過打亂LINC複合體及中心體改變導致基因組不穩定-30
圖16、KRT6-KRT14/V7通過打亂LINC複合體及中心體改變導致基因組不穩31
圖17、K6-K14 / V7通過基因表達去影響組蛋白活化---------------------------------32
圖18、K6-K14 / V7通過基因表達改變調節葡萄糖和酮體的代謝控制------------33
參考資料(References)--------------------------------------------------------------------------- 34

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