紅茶酒精萃取物對人類口腔癌細胞SCC-4侵襲轉移及蛋白酶表現影響之研究

近年來台灣地區十大癌症中，口腔癌的發生率和死亡率皆有逐年升高的趨勢，而癌細胞的轉移往往是臨床治療上最難以根治及導致死亡率增加的主因，根據統計約有90％為鱗狀細胞癌，其在口腔發生的部位以舌部最多，而且大概50％會發生頸部淋巴結轉移。紅茶，是世界廣為流傳的一種飲品，具有抗氧化的生物活性，但對於紅茶抑制癌細胞的侵襲和轉移作用以及其詳細機轉仍不明確。因此我們選用SCC-4這株高度惡化的人類舌鱗狀上皮癌細胞株來進行本次實驗。在本實驗中，利用50％的乙醇萃取紅茶之萃取物，再利用高效液相層析儀分析紅茶之萃取物，之後利用紅茶之萃取物處理SCC-4細胞株，然後進行MTT assay，發現紅茶之萃取物並不會影響SCC-4細胞的存活率；接著使用Transwell assay進行觀察並分析，發現紅茶之萃取物具有抑制SCC-4細胞的侵襲能力（p < 0.001）和運動能力（p < 0.01）；並且進行細胞-基質黏著分析，亦發現紅茶之萃取物對SCC-4細胞的黏附能力（p < 0.001）具有抑制效果；然後再以細胞延展分析進行實驗，發現紅茶之萃取物對SCC-4細胞的延展能力同樣具有抑制效果；而在明膠蛋白酵素電泳法、酪蛋白酵素電泳法以及冷光酵素測定中，同樣發現紅茶之萃取物對SCC-4細胞所分泌之MMP-2和u-PA蛋白酶活性表現以及基因啟動子轉錄活性具有顯著地抑制效果。綜合以上結果，紅茶之萃取物或許可應用在預防口腔癌的轉移或輔助口腔癌的治療上。

關鍵字

紅茶之萃取物；口腔癌；侵襲

並列摘要

The incidence and mortality of oral cancer in Taiwan have been constantly increased during the last decade and this increase in mortality could be mainly resulted from the difficulty in treatment caused by metastasis. More than 90% oral cncers have been identified as squamous epithelium cell carcinoma, especially in tongue. And then, nearly 50% of patients with oral squamous cell carcinoma (OSCC) present with pathologic or clinical evidence of lymph node metastasis. Black tea, the most widely consumed beverage in the world, has been evidenced to possess prominent antioxidant ability. However, the detailed effects and mechanisms of black tea on cancer cell invasion and metastasis were still unclear. Therefore, SCC-4 cells, a malignant oral squamous carcinoma cell line, was used in this study. First of all, SCC-4 cells were subjected to a treatment with 50% ethanol extracts of black tea leaves. Furthermore, the chemical profile of black tea polyphenol extracts (BTE) was analyzed by using a high-pressure liquid chromatograph (HPLC)−mass spectrometer (MS). And then MTT assays. Results suggested that black tea extract(BTE) have no effect on cell viability of SCC-4 cells. The Transwell assay revealed that a treatment of BTE significantly inhibited the cell invasion (p < 0.001) and motility (p < 0.01) capacities of SCC-4 cells. Next, we used the cell-matrix adhesion assay to determine the cell adhesion capacity. Here, we found the BTE exerted an inhibitory effect on cell adhesion (p < 0.001) of SCC-4 cells. Therefore, the results of the cell spreading assay also showed that the same inhibitory effect on cell spreading of SCC-4 cells. Zymography and promoter luciferase analysis revealed that BTE inhibited the proteinase and transcriptional activities of MMP-2 and u-PA of SCC-4 cells, respectively. These results suggested that BTE may be beneficial prevention of cancer metastasis, and helpful development of cancer therapy.

並列關鍵字

black tea extract ； oral cancer ； invasion ； proteinase

參考文獻

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紅茶酒精萃取物對人類口腔癌細胞SCC-4侵襲轉移及蛋白酶表現影響之研究

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