中心體蛋白CPAP (Centrosomal P4.1 associated protein) 除了在中心體功能中扮演重要的角色，同時也具有共同轉錄活化子(transcriptional co-activator)的角色。細胞介素6號(IL6)/訊息傳遞轉錄活化因子3號(STAT3)的活化路徑在許多癌細胞中被發現，包括肝癌。我們之前的研究發現，CPAP是STAT3的共同轉錄活化子，會透過與STAT3的交互作用，增加STAT3的轉錄活性以及其下游基因的表現。同時，CPAP亦參與肝癌的血管新生及惡化。此外，我們發現一個具有132個胺基酸的新穎胜肽N66，能阻隔CPAP與STAT3的相互作用，進而抑制STAT3的轉譯活性，達到阻斷肝癌中STAT3活化所驅動的癌細胞生長以及血管新生作用。本論文主要的研究目的在進一步找出N66抑制CPAP所導致的STAT3轉錄活性的最小有效區域，並且進行功能性試驗。透過免疫沉澱法、原位蛋白標定技術、即時聚合酶鏈式反應，以及細胞爬行等實驗，我們已經將N66的有效區域縮小到33個胺基酸 — N66-3。我們的實驗結果可確認N66-3在將來有機會發展成為在CPAP大量表現的肝癌中抑制IL6/STAT3路徑途徑之蛋白藥物。

In Taiwan, Hepatocellular carcinoma (HCC) is the second cancer-related death in Taiwan. CPAP (Centrosomal P4.1 associated protein) participates in activating interleukin 6 (IL6)/STAT3 pathway and contributes to tumorigenesis and angiogenesis in HCC. Our previous studies indicated that CPAP promotes the activation of STAT3 and its downstream effects in response to IL6 stimulus via forming complex with STAT3. We found a novel peptide N66 can decrease the STAT3-mediated angiogenesis and tumorigenesis in HCC. In this study, we aimed to map the smallest functional domain of N66 in blocking the CPAP-mediated STAT3 activation and investigated its effects in inhibiting tumorigenesis. Using colony formation, reporter assay, RT-qPCR, and migration assay, we mapped the smallest functional fragment of N66, named N66-3. Additionally, immunoprecipitation (IP) assay and in situ proximity ligation assay (PLA) both demonstrated the interactions between the N66-3 with STAT3. Our results suggest that the N66-3 peptide may serve as a potential protein drug to target IL-6/STAT3 in CPAP-overexpressing HCC.

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