因染色體轉位而產生的融合基因，是癌症基因體中最常出現的體變異形式，其中令人存疑的現象是，多於80%的融合基因是從癌症發生率只佔10%的血癌、淋巴癌、骨癌及軟組織肉瘤中發現；而佔癌症死亡率80%的上皮癌中卻只發現大約10%的融合基因。這樣的現象不禁讓人懷疑，是否有更多未知的融合基因存在於上皮癌中。另外，至今已經證實許多針對有酵素活性且會參與癌症發生的融合基因的抗癌藥物，可以有效的治療帶有這些融合基因的病患，更突顯了融合基因在癌症治療的重要性。因此這個研究的主要目的便為：建立一個高通量的檢驗方法，以適用於尋找上皮癌中潛藏的可用藥物治療的融合基因。
快速放大cDNA末端法(rapid amplification of cDNA ends, RACE)是一個常用於尋找融合基因的技術，不過它可能會因為細胞中fusion/wild-type transcript ratio比例很低，而必需進行大量DNA定序。因此我們在RACE之後接續進行基因表現連續分析法(serial analysis of gene expression, SAGE)以提高檢驗的通量，經由SAGE將DNA序列濃縮為可代表基因特性的短tag鏈後，可以將DNA定序的效率提高到20~30倍以上。我們這個將RACE和SAGE結合的檢測方法定名為：擴增後轉錄產物連續分析法(serial analysis of amplified transcripts, SAAT)。
為了建立以及評估SAAT的運作，我們利用一個已知有多達64種translocation partner genes的MLL (Mixed Lineage Leukaemia) 基因為模型。在這個研究中我們藉著MLL完善的基礎研究成功的建立了一個對MLL基因有專一性的3’-RACE，並接續著利用一株帶有MLL轉位基因的細胞株(MV 4-11)來評估SAAT的準確性及靈敏度。將來我們將匯集許多上皮癌病患的檢體，希望藉著單一次SAAT的進行便對全部的檢體進行偵測，以研究其中是否有潛藏的可用藥物治療的融合基因。

Fusion genes, resulting from chromosome translocations, are the most prevalent form of somatic changes in the cancer genome. Intriguingly, >80% of all known fusion genes are attributed to leukemias, lymphomas, but bone and soft tissue sarcomas that account for only 10% of all human cancers. In contrast, common epithelial cancers, which account for 80% of cancer-related deaths, can only be attributed to 10% of known recurrent fusion genes. Furthermore, if translocation-mediated fusion genes encode activated enzymes with direct oncogenic potential, targeting such enzymes could provide a feasible approach to treat individuals harboring the corresponding fusion genes. Therefore, a high-throughput method is needed not only for differential diagnosis but also for discovering drugable fusion genes in solid tumors.
RACE (rapid amplification of cDNA ends) is a widely used technique for finding fusion genes but its chief defect is the demand of a great deal of sequecing which may result from low fusion/wild-type transcript ratios. We propose to use SAGE (serial analysis of gene expression) after RACE to increase the throughput. When sequencing same amount of clones, SAGE can give 20 to 30 times more information than RACE alone. Therefore, it would be possible to pool the cases into groups and screened at once. This strategy is designated SAAT (serial analysis of amplified transcripts).
In order to assess the performance of SAAT we use MLL (Mixed Lineage Leukaemia) translocations as an example. MLL translocations occur in up to 70% of infant Acute lymphoblastic leukemia (ALL), and in about 3% of acute myeloid leukemia (AML), and it is already known can be recombined with 64 different partner genes, posing a challenge for routine molecular diagnosis.
In this research we based on the intergraty of the basic reaserch of MLL gene, sucessfully established a RACE system specific to MLL gene.Then we mixed normal leukocyte and MV4-11 that have MLL-AF4 translocation at different ratio to test the correctness and the detection efficiency of SAAT.In the future, we well use SAAT to screen a pool of samples from carcinoma patients at a time, for the purpose of discovering drugable fusion genes in solid tumors.

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