近年來的研究發現到在非小細胞肺癌病患的檢體中有越來越多新的融合基因被診斷出來，例如EML4-ALK融合基因佔肺小細胞肺癌約5~8%。於是本實驗室也想從非小細胞肺癌病患的檢體之中能否發現到新的融合基因。在實驗室先前的研究之中，我們透過多種分子檢測方式(IHC、FISH、RT-PCR、RACE)從100多例非小細胞肺癌病人的蠟塊檢體之中發現到1例新穎的ALK融合基因，5’端為Sperm Antigen With Calponin Homology And Coiled-Coil Domains 1-Like (SPECC1L) 與3’端為Anaplastic Lymphoma Kinase (ALK) 融合所形成的SPECC1L-ALK，所以本篇研究主要想探討新穎ALK融合基因是否在細胞株中與臨床所常見的EML4-ALK基因的生物特性是否相似，另外也會去比較新穎的融合基因SPECC1L-ALK和EML4-ALK融合基因在細胞株之中對於Tyrosine kinase inhibitor (TKI) 治療功效是否有所不同，由本篇實驗數據得知，新穎的融合基因SPECC1L-ALK和EML4-ALK皆能促進細胞增生，細胞侵襲/遷移和上皮-間質轉化(EMT)。在ALK TKI 處理48小時後進行WST-1和Colony formation實驗，研究得知，經過藥物處理後的ALK-rearranged cell的細胞存活率會降低。綜合上述研究結果，新穎的ALK融合基因SPECC1L-ALK生物特性與其ALK常見融合基因EML4-ALK相似，但在藥物敏感性實驗方面，HEK293-EML4-ALK比HEK293-SPECC1L-ALK稍敏感，至於此新穎的融合基因是否在臨床上有明顯的臨床意義，我們需要進一步去探討。

A fusion gene, echinoderm microtubule associated protein like 4 – anaplastic lymphoma kinase (EML4-ALK), with transforming activity has recently been identified in 5~8 % of non-small cell lung cancer (NSCLC). Previously in our laboratory, a novel ALK gene fusion partner, Sperm Antigen with Calponin Homology and Coiled-Coil Domains 1-Like (SPECC1L) was found in a patient with NSCLC through 5’-RACE, which was confirmed by RT-PCR, sequencing and FISH. The aim of this study is to establish SPECC1L-ALK as an oncogenic ALK aberration which is a candidate for targeted therapy. Stable cell pools expressing SPECC1L-ALK, EML4-ALK, or wild-type SPECC1L were prepared. Western blotting revealed that the corresponding gene products were at the predicted molecular weights. Next, the novel SPECC1L-ALK fusion gene was subjected to a series of experiments to determine its oncogenic characteristics, along with the well-known EML4-ALK fusion as a comparison. The results show that HEK293-SPECC1L-ALK and HEK293-EML4-ALK similarly increased the cell proliferation, cell migration/invasion, and epithelial mesenchymal transition (EMT) by activating the ALK downstream signaling pathway. There are several tyrosine kinase inhibitor (TKI) available for the treatment of NSCLC harboring ALK fusion genes, such as crizotinib. Their efficacies against SPECC1L-ALK and EML4-ALK were compared using a cellular model. The results revealed that crizotinib can inhibit the cell growth and cell survival in ALK-rearranged HEK293 cell.In summary, SPECCL-ALK was a novel ALK fusion gene similar to EML4-ALK. However, SPECC1L-ALK seemed to be less sensitive to crizotinib as compared to EML4-ALK in the HEK293 cell model. Its clinical significance needs further investigation.

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