評估川陳皮素在非小細胞肺癌中的治療潛力： Fascin-1在NSCLC發病機制中的作用

非小細胞肺癌 (non-small cell lung cancer, NSCLC) 為全球癌症相關死亡的主要原因之一，具高侵襲性與轉移潛能，因此開發更安全有效的治療模式為目前主要策略。Fascin-1作為一種肌動蛋白綑綁蛋白，其高表現除了與不良預後顯著相關，也在癌細胞遷移與上皮間質轉化 (epithelial-mesenchymal transition, EMT) 中扮演關鍵角色。透過生物資訊平台GEPIA (Gene Expression Profiling Interactive Analysis，網址：http://gepia.cancer-pku.cn/) 對TCGA (The Cancer Genome Atlas) 資料進行分析顯示NSCLC組織中fascin-1表現顯著高於正常肺組織，並且與病人預後不佳密切相關，具有成為潛在治療標靶的潛力。本研究聚焦於天然化合物川陳皮素 (nobiletin) 是否可藉由調節fascin-1表現來抑制NSCLC對細胞的侵襲性與遷移能力及分子機制。我們利用A549細胞株進行MTT細胞存活分析、傷口癒合、Transwell遷移與侵襲實驗，並透過西方墨點法 (Western blotting) 與螢光免疫染色 (Immunofluorescence staining) 進行蛋白表現及細胞骨架觀察等多項技術進行探討。 結果顯示，nobiletin可劑量依賴性地抑制fascin-1表現，並進一步抑制F-actin聚合與偽足形成，導致NSCLC細胞遷移與侵襲能力明顯下降。此外，nobiletin亦可抑制AKT Ser 473位點的磷酸化 (phosphorylated AKT, p-AKT)，並降低下游β-catenin與NF-κB p65之表現，推測fascin-1可能參與PI3K/AKT/β-catenin及NF-κB等訊號路徑之下游調控。細胞功能實驗亦證實nobiletin除了抑制A549細胞存活與傷口癒合能力，也誘導細胞形態由具侵襲性的紡錘狀轉變為上皮樣穩定型態，顯示其對EMT過程具干擾作用。總而言之，nobiletin藉由影響fascin-1表現，進而干擾相關訊號傳導與細胞骨架動態，展現其抑制NSCLC細胞侵襲、遷移與EMT的多重抗腫瘤效應。本研究首次揭示fascin-1可能作為nobiletin的靶點之一，強化其在腫瘤微環境與轉移機制中的關鍵角色，並為未來發展以fascin-1為核心的肺癌輔助治療策略提供重要實驗依據，有望成為未來的輔助治療選項，進一步應用於肺癌轉移抑制與精準醫療。

Non-small cell lung cancer (NSCLC) is one of the leading causes of cancer-related deaths worldwide, largely due to its aggressive nature and ability to spread. Fascin-1, an actin-bundling protein, is found to be highly expressed in NSCLC and is linked to poor prognosis, cancer cell movement, and epithelial-mesenchymal transition (EMT). Analysis of the Cancer Genome Atlas (TCGA) database using the bioinformatics tool GEPIA (Gene Expression Profiling Interactive Analysis, http://gepia.cancer-pku.cn/) showed that fascin-1 levels are significantly higher in NSCLC tissues compared to normal tissues, correlating with negative patient outcomes. The findings suggest that targeting fascin-1 could be a promising strategy for treating NSCLC.The potential of the natural compound nobiletin to diminish the translocation and invasion capabilities of NSCLC cells is explored in this research, focusing on the modulation of fascin-1 expression and its related molecular mechanisms. Using A549 cells, we performed MTT cell viability assay, wound-healing assay, and Transwell migration and invasion assays. Additionally, we assessed protein expression and cytoskeletal alterations through Western blotting and immunofluorescence staining.Our findings demonstrated that nobiletin reduced fascin-1 expression in a dose-dependent manner, inhibited F-actin polymerization and filopodia formation, thereby decreasing the invasion and migration of NSCLC cells. Nobiletin also inhibited AKT phosphorylation (p-AKT) and lowered the expression of downstream targets β-catenin and NF-κB p65, suggesting that fascin-1 may be involved in the downstream effects of the PI3K/AKT/β-catenin and NF-κB signaling pathways. Functional experiments further confirmed that nobiletin reduced A549 cell viability and wound-healing capacity, leading to a change in cell morphology from invasive spindle-like shapes to stable epithelial-like forms, indicating a suppression of the EMT process. In summary, nobiletin demonstrates various antitumor effects in NSCLC by regulating fascin-1 expression, disrupting related signaling pathways, and modifying cytoskeletal dynamics. This study is the first to identify fascin-1 as a potential target of nobiletin, highlighting its crucial role in the tumor microenvironment and metastatic processes. These results provide important evidence for the development of fascin-1-targeted adjuvant therapies for lung cancer, with potential implications for metastasis prevention and precision medicine.