探討印度紅茶及theaflavin 3,3\'-digallate透過調控上皮-間質轉換及蛋白酶抑制黑色素瘤細胞轉移之相關機轉研究

研究指出長期暴露在陽光下容易罹患皮膚癌，在早期皮膚癌通過手術就能有98%治療成功，但要治療轉移型皮膚癌相當困難。從古至今茶文化相當盛行，在茶類中有很多關於綠茶的研究，但是關於紅茶對人類黑素細胞瘤的作用的研究還很有限。本研究中以紅茶萃取物(black tea ethanol extract; BTEE)處理人類黑色素瘤A 375及A2058細胞作為研究，以MTT assay測定細胞活性試驗。Zymography探討matrix metalloproteinase-2 (MMP-2)、matrix metalloproteinase-9 (MMP-9)及urokinase-type plasminogen activator (u-PA)活性變化。傷口癒合試驗和Boyden chamber觀察細胞的移動和侵襲能力。細胞群落試驗看細胞群落生成。細胞貼附試驗觀察細胞貼附能力。西方墨點法觀察其細胞內訊息傳遞路徑。結果發現，BTEE能抑制細胞的MMPs、u-PA活性、細胞的爬行與侵襲能力及上皮間質轉換(epithelial-mesenchymal transition; EMT)路徑。因此接著加入促進劑腫瘤壞死因子(tumor necrosis factor-; TNF-)後發現，BTEE對TNF-所誘導黑色素細胞瘤之MMPs、u-PA表現量的影響。此外，利用紅茶純化物theaflavin 3,3'-digallate (TF3)也證實，TF3可有效抑制細胞的MMPs、u-PA與細胞爬行及侵襲能力。TF3也可降低TNF-所誘導黑色素細胞瘤之MMPs、u-PA表現量。

Studies have shown that prolonged exposure to the sun can lead to skin cancer. Surgical treatment of early-stage skin cancer has a 98% cure rate, but late-stage skin cancer is more difficult to cure. Since ancient times, tea culture has been quite popular. There are many studies on green tea among tea types, but the studies on the effect of black tea on human melanoma are still very limited. In this study, melanoma cells A 375 and A2058 were treated with black tea ethanol extract (BTEE). MTT assay was used to assay cell activity. Zymography was used to dertermine activities of matrix metalloproteinase (MMP)-2, MMP-9, and urokinase-type plasminogen activator (u-PA). The Wound-healing assay and Boyden Chamber invasion assay observed the migration and invasion ability of cells. Colony formation observed the formation of cell communities. The adhesion ability was observed by cell-matrix adhesion assay. Western blot were used to observe which pathway was inhibited. It was found that BTEE could inhibit the MMP and u-PA activity, effectively inhibit the migration and invasion of cells, and also inhibit the EMT pathway of cells. After adding the promoter tumor necrosis factor-(TNF-, it was found that BTEE had an inhibitory effect on the expression of MMPs and u-PA in melanoma cells induced by TNF-. Theaflavin 3,3'-digallate (TF3) was also found to inhibit MMP, u-PA, cell migration, and invasion. TF3 also reduced the level of MMPs and u-PA induced by TNF- in human melanoma cells.