| Summary: | 碩士 === 元培科技大學 === 醫學檢驗生物技術研究所 === 98 === Background: Cancer become the major leading cause of death in Taiwan during recent two decades and the strategies of prevention and treatment for cancer is more important. Many plant polyphenols have been demonstrated in vivo or in vitro anti-cancer potential, indicating the rich variety of fruit polyphenols exhibit great potential for development into anti-cancer drugs. Grape seed and longan seed have been shown rich in polyphenolic compounds. In this thesis, the polyphenols of these two fruit seeds were extracted and tested their anti-cancer activity.
Methods: Grape seed extract (GSE) and Longan seed extract (LSE) were measured the polyphenols by colorimetry. Lung (A-549), liver (Hep G2), cervical (C-33A), and breast (MDA-MB-231) cancer cell lines were treated with LSE and two pancreatic cancer cell lines (Paca-2 and Bxpc-3) were treated with GSP. The treated cells were respectively assessed for viability by trypan blue exclusion, for cell cycle distribution by flow cytometry, for apoptosis by annexin V labeling, for changes in the levels of proteins involved in cell cycle control or apoptosis by immunoblotting, and for cell invasion and metastasis by Borden and artificial basement membrane channel capacity.
Results: GSE and LSE potently show the proliferation and colony forming activity on the treated cancer cells. The cellular mechanisms include GSP inducing G1 phase arrest in Bxpc-3 and apoptosis in Paca-2. LSE induces G1 or S phase arrest in A-549, MDA-MB-231, and C-33A cells and apoptosis in Hep G2 and C-33A cells. Western blotting indicated that LSE and GSP mediated cell cycle arrest by inducing the overexpression of p21 and p27 and by attenuating the expression of cyclin D1 and cyclin A. GSE and LSE also lead to a reduction in protein levels of Bcl-2 and activated caspase 3 in apoptotic cells. GSE also inhibit invasion and metastasis of both Paca-2 and Bxpc-3 and by attenuating the expression of MMP-2 and MMP-9. The inhibitory effects of GSE and LSE on cell growth of different types of cancer cell lines are mainly from inducing cell apoptosis and/or cell cycle arrest. The results indicate that GSE and LSE is a potential chemopreventive and treatment agent for human cancer.
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