共病與癌症期別影響台灣黑色素瘤患者預後及欖仁葉萃取物對人類黑色素瘤細胞抗癌轉移之研究

背景 1 : 共病 (comorbidity) 與癌症期別可能會影響台灣的黑色素瘤的患者預後，相關研究在台灣從未涉及，需要進一步了解。 方法 1 : 我們利用台灣健保資料庫做了回顧性的研究， 從 2009 年到 2017 年的癌症登記檔，選取 476 位黑色素瘤患者當研究組，另外從 2000 年至 2018 年選取未被診斷黑色素瘤的族群當對照組，以查爾森共病指標 (charlson comorbidity index) 計算黑色素瘤患有共病的輕重，另以 cox proportional hazards model 估算疾病的風險比， 結果 1 : 黑色素瘤患者患有共病的比例佔有 55.5%， 共病的高盛行率與黑色素瘤的高期別具有相關性，較多共病的區域轉移及遠端轉移的患者有較高的死亡風險，在兩組的比較下，雖然都顯示共病越多，死亡率會隨著增加，但在黑色素瘤患者的死亡率更明顯高於未患黑色素瘤的組別。結論 1 : 高期別具有越多的共病與較高死亡率有顯著相關。背景 2 : 黑色素瘤是一種具有致命性，侵襲性的皮膚惡性癌症，初期發現，在患處予以切除即有滿意的療效，但在中後期的階段，使用手術，化療，免疫療法及放射線處置，都無法獲得有效的治療；黑色素瘤一旦進入轉移的階段，治療的效果就不如預期。 欖仁葉萃取物 (Terminalia catappa leaves extract：TCE) 含有大量的多酚及黃酮，具有掃除自由基，抗氧化及抗基因毒性的的作用。 方法 2 : 在A2058 和 A375 細胞實驗中，我們使用細胞傷口癒合復原分析，細胞侵襲性與移動性分析來了解欖仁葉萃取物是否具有抑制細胞爬行能力、侵襲性與移動性的效果；西方墨點法驗證欖仁葉萃取物是否抑制 -catenin、p-Src (Tyr 416)、Vimentin 與 p-FAK (Tyr397) 蛋白表達的功能，另外 gelatin zymography 及 casein zymography 則是測定欖仁葉萃取物是否分別抑制 MMP-2 及 u-PA 的活性。 結果 2 : 欖仁葉萃取物能顯著抑制 MMP-2 與 u-PA 的活性，兩株細胞的爬行能力，侵襲性與移動性也被明顯的抑制，西方墨點法也有效抑制 -catenin， p-Src (Tyr416)，另外在 A2058 的細胞裡也對 Vimentin 與p-FAK (Tyr397) 產生很好的抑制效果。 結論 2 : 總而言之，欖仁葉萃取物對轉移的黑色素瘤具有顯著的抑制效果，未來應用於黑色素瘤的癌轉移治療上可能是一種很有潛力發展的藥物。

Backgroud 1: Comorbidity and stage may affect the prognosis of melanoma patients in Taiwan. But no report paid an attention on this study. Methods 1 : We identified 476 melanoma patients and 4,760 members of the matched comparison cohort. Patients with a primary diagnosis of melanoma by the Taiwan Cancer Registry from 2009 to 2017 were recruited as the study population. The comparison group was never diagnosed with melanoma from 2000 to 2018. The Charlson comorbidity index was conducted to calculate the subjects’ disease severity. Results 1 : An association between poorer survival and comorbidity was verified in this study. We found that the level of comorbidity was strongly associated with mortality. Conclusion 1 : Advanced stage of melanoma patients with more comorbidities was significantly associated with the higher risk of mortality rate. Backgroud 2 : Melanoma is a highly aggressive, lethal, and malignant cancer. Once diagnosed early, it can be easily removed and cured with satisfaction. Although many methods such as surgery, chemotherapy, radiotherapy, and immunotherapy have been used to treat this disease at an advanced stage, the outcomes are poor. Terminalia catappa leaves have been shown to have various biological benefits, including antitumor activity. The specific effects and molecular mechanisms of Terminalia catappa leaf in treating A2058 and A375 melanoma cells in vitro need to be clarified. Methods 2 : Modified Boyden chamber assays were used to investigate whether Terminalia catappa leaf extracts inhibited A2058 and A375 cell migration/invasion capacities. The activities of matrix metalloproteinases-2 (MMP-2) and urinary type plasminogen activator (u-PA) were clarified through the zymography assay. Using Western blot to assess that whether TCE reduced the expression of phospho-Src (Tyr416), phospho-focal adhesion kinase (FAK) (Tyr397), Vimentin, and β-catenin. Results 2 : Modified Boyden chamber assays demonstrated that Terminalia catappa leaf extracts treatment significantly inhibited A2058 and A375 cell migration/invasion capacities. In the zymography results, we showed that Terminalia catappa leaf extracts (TCE) negatively modulated the activities of matrix metalloproteinases-2 (MMP-2) and urinary type plasminogen activator (u-PA). Western blot indicated that TCE reduced the expression of phospho-Src (Tyr416), phospho-focal adhesion kinase (FAK) (Tyr397), Vimentin, and β-catenin. Conclusion 2 : In this study, TCE affected the metastasis of the A2058 and A375 melanoma cell lines by inhibiting the FAK/Src interaction and Wingless-int1 (Wnt)/β-catenin pathways in vitro. TCE may serve as a powerful chemopreventive agent against metastasis of melanoma cancer.