李健群黃雅莉Ya-Li Huang營養學研究所2026-03-249999/12/312015https://handle.ncl.edu.tw/11296/427t52https://scholars.csmu.edu.tw/handle/123456789/6997乳癌為全球女性常見惡性腫瘤，其癌細胞高度轉移力是造成乳癌患者預後不佳而導致死亡的主因。Twist1為一具有basic helix–loop–helix (bHLH)結構之轉錄因子，近期研究證實，Twist1在促使癌細胞轉移、血管新生及上皮間質轉化 (Epithelial-mesenchymal transition, EMT)過程中扮演重要角色。另有文獻指出，Wnt-1/β-catenin訊號路徑異常活化與促使STAT3α表現及活化進而增加Twist1表現有關。本實驗室已成功建立細胞癌化促進劑12-O-tetradecanoylphorbol-13-acetate (TPA)誘導MCF-7人類乳癌細胞轉移模式，本研究續以此模式釐清Twist1在MCF-7細胞轉移扮演之角色及相關調控路徑，同時觀察二十二碳六稀酸 (Docosahexaenic Acid; DHA)是否透過影響Twist1表現而抑制TPA誘導之癌細胞轉移。研究結果顯示，給予不同劑量TPA (0, 1, 10, 100 ng/ml)處理細胞24小時後可顯著誘發p-GSK-3β、Wnt-1蛋白表現及STAT3、β-catenin、Twist1蛋白和mRNA表現。分別預處理JNK抑制劑-SP600125、ERK抑制劑-PD98059、p38抑制劑-SB203580、PI3K抑制劑-LY294002及STAT3抑制劑-WP1066 1小時後再添加TPA，觀察不同抑制劑對TPA誘發Twist1表現之影響，結果顯示僅有WP1066顯著抑制Twist1表現。預處理DHA可降低TPA誘發p-GSK-3β、STAT3、β-catenin、Twist1蛋白表現。此外，利用siRNA技術將Twist1 knockdown後可延遲TPA所誘發癌細胞移行。傷口癒合試驗及觀察Twist1在TPA誘發癌細胞移行之能力。結論：TPA可透過增加Wnt-1及活化GSK-3β而增加β-catenin、STAT3表現，最終促進Twist1表現，使腫瘤細胞產生轉移，而DHA可透過抑制TPA所誘發Wnt-1/β-catenin/STAT3訊號路徑，抑制Twist1表現進而降低MCF-7乳癌細胞移行。Breast cancer is the most common cancer in women worldwide. Tumor metastasis is a leading major cause of cancer death among breast cancer patients. Twist1 is a basic helix loop helix (bHLH) transcription factor essential in embryological morphogenesis. Previous studies have demonstrated that Twist1 plays an important role in cancer metastasis,angiogenesis and epithelial-mesenchymal transition (EMT). Abnormal activation of Wnt-1/β-catenin signaling pathway was found to induce the expression of STAT3α, and subsequently results in tumorigenesis and metastasis. In this study, tumor-promoting agent-12-O-tetradecanoylphorbol-13-acetate (TPA) was used to induce MCF-7 cell metastasis, and clarify the role of Twist1 involved in tumor metastasis. Furthermore, we investigated whether the anti-metastasis potential related to down-regulation of Twist1 expression by DHA in TPA-induced MCF-7 breast cancer cells and the possible mechanisms involved. We found that TPA significantly induced Twist1, Wnt-1, β-catenin, p-GSK-3β, and STAT3α protein expression and 100 μM DHA significantly inhibited TPA-induced p-GSK-3β, β-catenin, STAT3α, Twist1 in MCF-7 cells. Wound-Healing assay showed that TPA-induced MCF-7 cell migration was suppressed by Twist1 siRNA. Taken together, these results suggested that TPA-induced cell migration is associated with activation of Wnt-1/β-catenin and GSK-3β signaling pathway, leading to STAT3α and Twist1expression. Attenuation of TPA-induced Wnt-1/β-catenin/STAT3Twist1Wnt-1/β-cateninSTAT3αMCF-7Migration乳癌thesis