研究葡萄籽萃取物對人類非小細胞肺癌細胞之細胞凋亡的影響

The Investigation of Grape Seed Extract’s Effects on Proliferation and Cell Apoptosis of Non-Small-Cell Lung Carcinoma Cell Lines

尤貞惠(Chen-Hui Yu)

指導教授 ： 陳炳宏

高雄醫學大學/生命科學院/生物科技學系碩士班/碩士(2016年)

摘要

葡萄籽萃取物（GSE）是目前的研究指出具有抗氧化及抗癌能力的物質，而肺癌是我國最常見的癌症，因此我們想了解GSE是否能抑制肺癌。我們用了兩株非小細胞肺癌細胞，H1299和A549觀察GSE所造成的影響，結果顯示，GSE能夠抑制非小細胞肺癌細胞H1299及A549的生長並促進這兩株細胞的死亡，所以我們進一步探討GSE是否能對肺癌細胞產生細胞凋亡。實驗結果顯示GSE能夠對肺癌細胞誘發細胞凋亡。當我們確認了GSE能夠造成細胞凋亡，我們想進一步了解GSE造成細胞凋亡的關鍵步驟。實驗證明GSE誘發細胞內活性氧物質的產生、活化與粒線體相關的p38及SAPK/JNK訊息傳路徑和降低粒線體膜電位。另外，我們發現GSE引發內質網壓力並增加了和內質網壓力有關的細胞凋亡蛋白質CHOP的表現。實驗也顯示GSE造成NSCLC細胞的雙股DNA斷裂而造成細胞凋亡。為了知道GSE是否會對正常細胞造成影響，我們使用永生型人類肺部上皮細胞BEAS-2B作為正常細胞的模型。實驗結果顯示，GSE不會增加BEAS-2B細胞的細胞內活性氧物質、誘發細胞凋亡蛋白質的表現及造成雙股DNA斷裂。因此，我們推斷GSE不會影響正常細胞並具有抑制非小細胞肺癌的潛力。

關鍵字

葡萄籽萃取物 ； 非小細胞肺癌 ； 內質網壓力 ； 雙股DNA斷裂 ； 細胞凋亡

並列摘要

Grape seed extract (GSE) has been researched as an anti-cancer and anti-oxidation substance. Lung cancer is the most common cancer in Taiwan. Therefore, we wondered if GSE can inhibit lung cancer. We used two different non-small-cell lung cancer (NSCLC) cell lines, A549 and H1299, to observe the effect of GSE treatments. The results showed that GSE can inhibit growth and induce cell death. Because of the induction of cell death, we tested the cell apoptosis of cancer cell lines after GSE treatments. The results revealed that, GSE induced apoptosis both in H1299 and A549 cells. Hence, we observed the protein expression of apoptosis. The result showed that GSE can induce protein expression of apoptosis. After we confirmed the apoptosis of GSE treatment, we asked what the key step of inducing apoptosis is. The experiments evaluated GSE can induced the intracellular reactive oxygen species (ROS), active p38 and SAPK/JNK, the mitochondrial-mediated signaling pathways, and decrease mitochondrial membrane potential. Besides, we found out that GSE induced the ER stress and increased CHOP expression, the protein related to ER stress and apoptosis. The experiments also showed that, GSE caused apoptosis by inducing DNA double-strand breaks (DSB) of NSCLC cells. To know that GSE if influence the normal cells or not. We used human lung/bronchus epithelial cell line, BEAS-2B, as the normal-like cell line. The results showed that, GSE didn’t increase the intracellular ROS, induce protein expression of apoptosis and DSB. Thus, we speculated that GSE doesn’t influence the normal cells and has potential for resisting non-small-cell lung cancer.

並列關鍵字

Grape seed extract ； Non-small-cell lung cancer ； ER stress ； DNA double-strand breaks ； apoptosis

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