丹酚酸A抑制人類鼻咽癌細胞轉移和侵襲之機制探討

The mechanisms of the inhibitory effects of Salvianolic acid A on migration and invasion of nasopharyngeal carcinoma cells

蔡明倢(Ming-Chieh Tsai)

指導教授 ： 楊順發

中山醫學大學/醫學院/醫學研究所/碩士(2020年)

摘要

鼻咽癌 (Nasopharyngeal carcinoma，NPC)是一種發生於鼻咽腔或上咽喉部的癌症，屬於頭頸癌的一種，雖然發生率低，但因不易檢查，早期症狀也不明顯，容易被忽略。基質金屬蛋白酶 (Matrix metalloproteinase，MMP)，為一個大家族，主要功能有分解細胞外基質，調控生長因子及細胞激素等，其中分解細胞外基質在癌細胞轉移中扮演重要角色。丹酚酸A (Salvianolic acid A)，為丹參萃取物之一，近年來發現有抑制MMPs以及抗癌的效果。然而目前丹酚酸A對鼻咽癌的影響尚未釐清。因此在本篇研究中將探討丹酚酸A在鼻咽癌中是否能抑制鼻咽癌細胞的轉移和侵襲能力。首先使用MTT assay 來觀察丹酚酸A對鼻咽癌細胞存活的影響，發現丹酚酸A不會影響細胞存活率。接著使用wound healing及boyden chamber實驗發現丹酚酸A皆會抑制鼻咽癌細胞的爬行及侵襲能力。再利用Gelatin zymography assay實驗觀察到丹酚酸A抑制鼻咽癌細胞分泌到細胞外的MMP-2活性，另外，在蛋白表現和mRNA表現方面也觀察到有相同的抑制效果。接著進一步用西方點墨法確認鼻咽癌細胞內的訊號傳遞路徑，發現丹酚酸A抑制Focal adhesion kinase (FAK)，Steroid receptor coactivator (Src)， Extracellular signal-regulated kinase(ERK)蛋白表現。綜合以上實驗，在不影響細胞存活率下，丹酚酸A透過ERK路徑抑制MMP-2 mRNA、蛋白表現與活性，進而抑制鼻咽癌細胞轉移能力。

關鍵字

鼻咽癌 ； 基質金屬蛋白水解 #37238 ； 丹酚酸A

並列摘要

Nasopharyngeal carcinoma (Nasopharyngeal carcinoma, NPC)is a type of cancer that occurs in the nasopharynx or upper laryngospasm. Matrix metalloproteinase (MMP)is a large family and its main function is to decompose the extracellular matrix, regulate growth factors and cytokines, among which the decomposition of extracellular matrix plays an important role in the metastasis of cancer cells. Salvianolic acid A (Sal A)is one of Salvia extracts. In recent years, it has been found to have the effect of inhibiting MMPs and anti-cancer. However, the effect of Sal A on nasopharyngeal cancer has not yet been clarified. Therefore, in this study we will investigate whether Sal A can inhibit the metastasis and invasion of nasopharyngeal carcinoma cells in nasopharyngeal carcinoma. Firstly, MTT assay was used to observe the effect of Sal A on the survival of nasopharyngeal carcinoma cells. It was found that Sal A did not affect the cell survival rate. Then we used wound healing and boyden chamber experiments to find that Sal A inhibits the ability of nasopharyngeal cancer cells to crawl and invade. Using Gelatin zymography Assay experiments, it was observed that Sal A inhibited the extracellular MMP-2 activity secreted by nasopharyngeal carcinoma cells. In addition, similar inhibitory effects were observed in protein expression and mRNA expression. Then, Western blotting was used to confirm the signal transmission pathway in nasopharyngeal carcinoma cells. It was found that Sal A inhibited the expression of Focal adhesion kinase (FAK), Steroid receptor coactivator (Src), Extracellular signal-regulated kinase(ERK) proteins. In conclusion, Sal A inhibits the expression and activity of MMP-2 mRNA and protein through the ERK pathway, thereby inhibiting the metastasis capacity of nasopharyngeal carcinoma cells.

並列關鍵字

Nasopharyngeal carcinoma ； matrix metalloproteinases ； Salvianolic acid A

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