2018-08-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/694378摘要：神經纖維瘤第一型(NF1)是常見的遺傳性神經外胚層疾病。大部分的神經纖維瘤(包括皮膚或是叢狀神經纖維瘤)是良性的；然而，大約有5~13%的病患，會從叢狀神經纖維瘤產生惡性神經纖維肉瘤(MPNST)。MPNST 是一種致命性的惡性腫瘤，其五年存活率不到20%。到目前為止，只有早期全部切除才能治癒MPNST；然而，MPNST 通常體積很大、容易復發及遠處轉移、有時解剖位置不容易切除等等原因，造成手術失敗情形相當常見。再者，放射治療以及化學治療的治癒率也不高。因此，尋找一個新的治療藥物，對於MPNST 的病患相當重要。在先前的研究中，薑黃素對於MPNST 有相當程度的抑制效果。我們使用薑黃素的萃取物Calebin-A 做初步的測試發現，Calebin-A 對於MPNST 細胞株有相當程度的抑制效果，而且其濃度明顯小於薑黃素。根據上述發現我們提出這個為期三年的研究，旨在確認Calebin-A在小鼠體內的功效，並了解殺傷腫瘤細胞的表觀遺傳機制（epigenetics），並進一步探討，處理後MPNST 分泌exosome 的內容物的變化。在第一年我們將使用Calebin-A 治療NF１病患切下來的組織所培養出來的細胞，緊接著是使用Calebin-A 來治療MPNST 異種移植小鼠的動物模式。為了解Calebin-A 治療對於MPNST 的效果是否和hTERT 及 COX-2 基因的表現相關，而這些表現量的差異是否透過表觀遺傳機制調控？我們第二年的計畫將研究在這兩個基因的histone acetylation, deacetylation 及CpG island 甲基化程度。最後，我們將會評估治療前後MPNST 分泌的exosomes 的內容物的變化，這些內容物包括：microRNA、soluble AXL 以及integrins 等等。在收取MPNST 培養液經萃取exosomes 之後，我們將使用次世代定序來評估microRNA 的變化。另外，soluble AXL 的表現量可以用ELISA 來定量，而我們將利用integrins cell adhesion array 的表現來評估哪一種integrin 在Calebin-A 治療之後有明顯改變。這些研究結果將非常有助於我們了解腫瘤發生後Calebin-A 治療的分子機制及其變化。<br> Abstract: Neurofibromatosis type 1 (NF1) is one of the most common hereditary neurological disorders. Most ofthe neurofibromas (cutaneous or plexiform NF) are benign; however, the life risk to develop a malignantperipheral nerve sheath tumor (MPNST), a deadly disease, is 5~13%. The 5-year survival rate forMPNST is less than 20%. Up to date, only radical resection can cure MPNST; but the large size withhigh recurrence, easily distant metastasis, and anatomical locations frequently make the surgeryunsuccessful. Radiotherapy and chemotherapy were failed in many trials. Thus, to identify anarmamentarium with easily deliverance and high bioavailability is urgently needed.In previous reports, curcumin had moderate tumoricidal effect of MPNST. We have recently obtained anew compound, Calebin-A which was extracted from curcuminoid. Our preliminary result demonstratedthe efficacy of Calebin-A in treating MPNST cell lines. We proposed this three-year study with aims toconfirm the efficacy of Calebin-A in vivo, to decipher the epigenetic mechanism of the tumoricidal effectand to further explore the changes in content of exosomes after treatment. In the first year, the replicationof efficacy in the primary cultured cells, neurofibroma and MPNST ex vivo will be carried out. Theeffectiveness would be further confirmed using a xenograft mouse model with oral feeding of Calebin-A.To uncover the epigenetic mechanisms of two MPNST related genes, hTERT and COX-2 after treatment,will be carried out in the second year. The histone acetylation including the activity of histone acetylaseand deacetylase, and changes of methylation of the CpG islands of promoter regions will be performed aswell. Furthermore, we would explore the changes of content of exosomes released by MPNST cells upontreating with Calebin-A in the third year. The exosomes will be purified from both the medium in vitroand the patients’ plasma in vivo. The changes of the sAXL, integrin species and microRNA profiles preandpost-treatment would be evaluated. These findings will be very helpful for us to understand themolecular mechanisms of tumorigenesis and the alterations after Calebin-A treatment.