根據臺灣國民健康署統計資料顯示，轉移性大腸直腸癌已多年蟬聯十大死因前三。我們目標蛋白IRSp53已知可調控細胞骨架重組進而促進偽足的形成。透過資料庫的分析，我們觀察到了高表達異構物IRSp53-T於大腸直腸癌與低存活率相關。進一步使用西方點墨法(Western blot)分析，晚期的大腸直腸癌擁有較多的IRSp53-T表達量比之早期。因而我們推測IRSp53-T可能涉及大腸直腸癌惡化的過程。本篇論文旨在探究IRSp53-T 於癌細胞中的轉移潛能，研究方法為透過建立穩定過表達IRSp53-T於大腸直腸癌細胞株SW480，該穩定過表達細胞株後續以53T代稱。細胞遷移以及侵襲試驗中，IRSp53-T展現了顯著的促進效應。而在轉移以及皮下小鼠模式中，我們於53T的組別中觀察到了巨噬細胞浸潤的團簇，然而其他異構物IRSp53-M和IRSp53-S並無類似的現象。西方點墨法中，IRSp53-T被觀察到促進了AKT的活化，而活化的現象推測與KRAS表現無關。確立轉移關係後，我們透過蛋白交互作用資料庫STRING以及現有文獻資料以釐清IRSp53-T與轉移相關的背後機制，最終選定IGF-1用於探測。相較於控制組別，53T細胞於IGF-1處置下展現持續性活化的AKT。鑒於AKT的活化位置在細胞膜周邊，我們使用免疫螢光染色偵測IGF-1處置後IRSp53-T在細胞中的分布位置。免疫螢光結果顯示IRSp53-T暴露於IGF-1下傾向於細胞膜周邊聚集。綜上所述，此研究結果顯示IRSp53-T具有促進大腸直腸癌轉移的潛力，並且我們推測IRSp53-T可能參與在IGF-1/AKT的路徑當中。

Metastatic colorectal cancer (mCRC) contributes a high mortality rate in Taiwan. IRSp53 has been known to regulate actin cytoskeleton organization and promote pseudopodia/lamellipodia formation. Through online database search, we observed that high expression of IRSp53-T was associated with poor survival in CRC. Western blot analysis also revealed higher levels of IRSp53-T in late stage than in early stage. These results suggested IRSp53-T may be involved in CRC progression. To characterize metastatic potential of IRSp53-T in vitro and in vivo, we generated a cell line constitutively expressing IRSp53-T in SW480 cells (abbreviated as 53T). Both transwell assay and invasion assay showed that IRSp53-T increased cell motility and invasion. In intraperitoneal metastatic and subcutaneous mouse model, we observed macrophage-infiltrating tumor nests in 53T, while recruitment of macrophages was not found in M and S isoforms. Western blotting analysis illustrated that IRSp53-T increased activation of AKT, which was independent of of KRAS levels. We used STRING database to acquire the potential pathway connecting IRSp53-T and metastasis and proposed the evidence-supported growth factor, IGF-1, as a candidate target. The results of western blot displayed constitutive activation of AKT in 53T cells compared with the control groups under IGF-1 treatment. Given AKT is activated in cell membrane, we further evaluated membrane distribution of IRSp53-T by immunofluorescent staining, which showed the clustering of IRSp53-T at cell membrane when exposed to IGF-1. Collectively, our results demonstrated the metastatic potential of IRSp53-T in CRC and possible involvement of IGF-1/AKT pathway.

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