抗菌胜肽Tilapia Hepcidin 1-5對抑制癌症細胞之研究

Study on inhibitory effects of Tilapia Hepcidin 1-5 in cancer cells

張婉婷(Wang-Ting Chang)

指導教授 ： 鄭鈞文 ； 陳志毅

中山醫學大學/醫學院/生化暨生物科技研究所/碩士(2010年)

https://doi.org/10.6834/CSMU.2010.00084

摘要

TH1-5 是來自於吳郭魚 (Oreochromis mossambicus) hepcidin 的一段抗菌胜肽。由結果顯示，TH1-5 這一個由22 個胺基酸組成的抗菌胜肽，會特異性的抑制人類纖維肉瘤細胞株HT1080 和人類子宮頸上皮癌細胞株 HeLa 的增生和移動，因此將對降低HT1080 細胞和
HeLa cell 的生長加以研究。利用MTT assasy、AO/EtBr 染色和soft agar assay 的實驗結果顯示，TH1-5 會抑制HT1080 細胞和HeLa 細胞的生長。而以SEM 和TEM 觀察細胞的形態，TH1-5 會將HT1080 細胞和HeLa 細胞之細胞膜破壞和摧毀。在wound healing assay 實驗顯示TH1-5 亦可抑制HT1080 細胞和HeLa 細胞的移動。以流式細胞儀分析，TH1-5 會誘導HT1080 細胞和HeLa 走向細胞凋亡並使細胞停滯在S 期。此外，在real-time PCR 結果，TH 1-5 處理HeLa 細胞會使Bcl-2、Bcl-xL，JNK, IL-6, IL-8, IL-10, IL-12, IL-13, IL-15, CAPN5,CTSG, p53, caspase-3, NFκΒ 和 INF-γ 之 mRNA 相較於沒有處理會高度表現，而在HT1080 細胞會使IL-2, CTSG 和INF-γ 高度表現。在由以上結果顯示TH1-5 對細胞的毒性的機制，將來可成為對抗人類纖維肉瘤和人類子宮頸上皮癌之化學治療藥物試劑。

關鍵字

癌症 ； 抗菌胜肽

並列摘要

TH1-5 from antimicrobial peptides of marine organisms, tilapia (Oreochromis mossambicus) hepcidin. The results indicated that TH1-5, a synthetic 22-mer antimicrobial peptide, specifically inhibited human fibrosarcoma cell (HT1080 cell line) and human cervical epithelioid
carcinoma cells (HeLa cell line) proliferation and migration. The way in which TH1-5 inhibited HT1080 and HeLa cell growth was then studied.TH1-5 inhibited HT1080 cell and HeLa cell growth in a concentration-dependent manner according to an MTT analysis, which was confirmed by a soft-agar assay and AO/EtBr staining. Scanning electron microscopy and Transmission Electron Microscope revealed that TH1-5 caused lethal membrane disruption in HT1080 cancer cells and HeLa cells, and a wound healing assay supported that TH1-5 decreased the migration of HT1080 cells and HeLa cells. A flow cytometric analysis,
showed that the TH1-5 induced apoptosis. In addition, Bcl-xL，JNK, IL-6,IL-8, IL-10, IL-12, IL-13, IL-15, CAPN5, CTSG and INF-γ mRNA expression was upregulated after treatment with TH 1-5 in HeLa cell to compared the untreated group. And in HT1080 cell, IL-2, IL-8, CTSG and INF-γ mRNA expression was upregulated. These findings suggest a
mechanism of cytotoxic action of TH1-5 and indicate that TH1-5 may be a promising chemotherapeutic agent against human fibrosarcoma cells and human cervical epithelioid carcinoma cells.

並列關鍵字

cancer ； antimicrobial peptide

參考文獻

Andrews, NC. Anemia of inflammation: the cytokine-hepcidin link.J.Clin. Invest. 2004; 113, 1251–1253.

Ashkenzi A, Dixit, VM. Death receptor: signaling and modulation.Science 218. 1998; 1305-1308.

cationic antimicrobial peptides. BBA Biomembranes.

Bortner CD, Cidlowski JA. Celluar mechanisms for the repression of

Impact of LL-37 on anti-infective immunity. J. Leukoc. Biol. 2005;