2010-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/649532摘要：肝細胞癌（簡稱肝癌）是全世界常見的癌之一，自從1984 至今，肝癌就高居台灣十大癌症死因的首位，每年大約有8000 國人死於肝癌，雖然目前以超音波及甲種胎兒蛋白定期追蹤檢查，可以早期篩檢出肝癌，然而肝癌的治療仍不十分的理想。肝癌治療的成效不佳的原因之一是發現太晚。雖然胎兒蛋白及DCP 都被用來做來診斷肝癌的腫瘤標記，可惜的是大約在1/3 的小型肝癌病人，其血中的胎兒蛋白仍是正常的，因此用胎兒蛋白及DCP 來早期診斷肝癌仍然不足，所以開發肝癌新的腫瘤標記仍然是需要的。最近的研究顯示，蛋白質的醣化現象與癌症的發生有關。醣化作用是蛋白質修飾的一個相當普遍的現象，蛋白質的醣化作用與細胞的生理功能有很大的關係。由於醣蛋白相對穩定，因此醣蛋白相當適合作為腫瘤的生物標記及作為腫瘤治療的標靶。在此計畫中，我們將利用蛋白質體學的方式來尋找與肝癌相關的醣蛋白。我們將收集肝癌開刀的病人，開刀前與開刀後的血清，並以有慢性病毒性肝炎但無肝癌的病人血清，及正常人的血清做為對照組。我們將以lectin 親和層析法來分離血清中的醣蛋白，然後用二次元電泳將蛋白質分離，以半定量的方式，來比較肝癌開刀的病人與對照組之間，有不同表現量的醣蛋白。根據分析的結果，選出有興趣的點，再用質譜儀或胺基酸定序儀分析其胺基酸序列，即可判別原蛋白質的身分。找出可能的肝癌相關的醣蛋白後，我們將以蛋白質晶片來確認這些醣蛋白後可以真的作為肝癌的腫瘤標記，我們也將利用免疫組織染色法，來看肝癌組織是否會表現我們所找到的這些醣蛋白。我們希望經由這個計畫，能找到可以早期診斷肝癌的腫瘤標記，進而提升肝癌治療的成績。<br> Abstract: Hepatocellular carcinoma (HCC) has been the leading cause of cancer death in Taiwan.Around 8000 people died of this cancer every year in Taiwan. Though regular sonographicexamination can early detect small HCC and there are many therapeutic modalities for HCC,the therapeutic results remains unsatisfactory. Though Alpha-fetoprotein (AFP) anddes-γ-carboxy prothrombin (DCP) are used as the tumor markers for diagnosis of HCCs, AFPis normal in around one third of small (< 3 cm) HCC patients. Elevated DCP activities werepresent in 44%-47.6% with HCCs less than 3 cm. Thus, these two markers are not goodenough for the early detection of small HCCs. To improve the survival, further investigationsof the early diagnostic markers are still needed.The development of HCC might be associated with glycosylation of many biomolecules.The AFP-L3 is one the examples. Glycosylation, one of the most ubiquitous forms ofposttranslational modification, plays important roles in a diverse set of biological processes. Ithas been shown that glycosylation is involved in signaling pathways associated with thetransformation of a normal cell to a cancer cell. Targeting at the glycoproteins couldpotentially harness the cancers. Thus, glycoproteins offer many advantages for potentialbiomarkers such as increased stability and solubility relative to unmodified proteins so thatissues around plasma collection and storage could be minimized.In this current project, we will apply the proteomic methods to identify theHCC-associated glycoproteins. Lectin affinity column will be used to isolate theglycoproteins. The glycoproteins will be separated by 2D-PAGE and semiquantitation will bedone to identify the glycoproteins. The protein spots of interest will be sequenced and proteinID could be obtained. Validation of the potential glycoprotein biomarkers will be done byprotein array on sera and by immunohistochemical stains on liver tissues. We believe that wecan find some glycoproteins associated with HCC. The glycoproteins could be potentiallyused as the early diagnostic markers.肝細胞癌﹐醣蛋白﹐腫瘤標記﹐蛋白質體學﹐胎兒蛋白hepatocellular carcinomaglycoproteinproteomicsbiomarkersAFP