- 學位論文
二甲雙胍抑制胃癌細胞生長之分子機制探討
Molecular Mechanism of Anti-diabetic Drug Metformin in Gastric Cancer Cell Growth
摘要
二甲雙胍(metformin)是目前廣泛使用於治療第二型糖尿病的用藥。其降血糖機制包括抑制肝臟葡萄糖新生作用、增加肌肉對葡萄糖的攝取、促進對胰島素的敏銳度。近期研究認為 metformin 或許會降低癌症發生率，但其抑制癌症的機制還不清楚，特別是在抑制胃癌的部分。在本研究中我們將評估 metformin 抑制人類胃癌細胞(AGS cells)的能力並利用相對和絕對定量的等量異位標籤(isobaric Tagging for Relative and Absolute Quantitation)定量蛋白質體學進一步探討抑制癌症的機制。我們的研究發現metformin抑制胃癌細胞增生的能力會隨著劑量增加而增加，metformin 也會誘發胃癌細胞G0/G1週期停滯，抑制胃癌細胞的細胞移動及影響胃癌細胞的細胞骨架之分布。我們利用定量蛋白質體學技術鑑定並定量177個蛋白質，並依照他們在控制組與metformin處理組的相對表現量挑選出9個表現量有差異的蛋白質，包含4個表現量上升的蛋白質(Prohibitin-2, Heterogeneous nuclear ribonucleoprotein A/B (hnRNP A/B), Clathrin heavy chain 1, Macrophage migration inhibitory factor (MIF))及5個表現量下降的蛋白質(T-complex protein 1 subunit epsilon, 26S proteasome non-ATPase regulatory subunit 2 (PSMD2), Stress-induced-phosphoprotein 1 (STIP1), Polypyrimidine tract-binding protein 1, Adenylyl cyclase-associated protein 1 (CAP1)). 其中蛋白質 STIP1 、 CAP1 及 PSMD2 在先前的研究中被指出與癌細胞的細胞增生和移動具有相關性。因此我們利用西方墨點法驗證 STIP1 、 CAP1 及 PSMD2 蛋白質的表現量並發現蛋白質表現趨勢與蛋白質體的分析結果是一致的。這些結果意味著 metformin 可能藉由調控胃癌細胞中蛋白質 STIP1 、 CAP1 及 PSMD2 的表現去抑制細胞增生和細胞移動。雖然還需更多的動物實驗或臨床實驗來驗證 metformin 的抗癌效果，本研究或許能提供胃癌治療的發展一個新的方向。
並列摘要
Metformin is one of the most widely used anti-diabetic drugs in type II diabetes treatment. The mechanism of lowering blood glucose is believed to be involved in suppressing hepatic gluconeogenesis, increasing muscular glucose uptake and enhancing insulin sensitivity. Recent studies suggest that metformin may reduce cancer risk, but the mechanism of anticancer action remains unclear, especially in gastric cancer. In our studies, we aim to evaluate the anticancer effects of metformin on human gastric cancer AGS cells, and use isobaric Tagging for Relative and Absolute Quantitation (iTRAQ)-based quantitative proteomics to further study the anticancer mechanism. Our results showed that metformin inhibited AGS cell proliferation in a dose dependent manner, induced AGS cell cycle arrest at G0/G1 phase, suppressed cell migration ability, and affected the distribution of cytoskeleton. Using quantitative proteomics approach, we identified 177 differentially expressed proteins upon metformin treatment; among these, nine proteins are significantly altered in expression level, including four up-regulated proteins (Prohibitin-2, Heterogeneous nuclear ribonucleoprotein A/B (hnRNP A/B), Clathrin heavy chain 1, Macrophage migration inhibitory factor (MIF)) and five down-regulated proteins (T-complex protein 1 subunit epsilon, 26S proteasome non-ATPase regulatory subunit 2 (PSMD2), Stress-induced-phosphoprotein 1 (STIP1), Polypyrimidine tract-binding protein 1, Adenylyl cyclase-associated protein 1 (CAP1)). In previous study, STIP1, CAP1 and PSMD2 were reported associated with cancer cell proliferation and motility. The expression level of STIP1, CAP1 and PSMD2 were further validated by immunoblotting and consistent with the proteomic results. Our findings imply that metformin might inhibit cell proliferation and migration in AGS cells by suppressing STIP1, CAP1 and PSMD2. Although more animal and clinical studies are needed to confirm the effects of metformin, our studies may provide a new way for developing therapies for gastric cancer.
參考文獻
延伸閱讀
- 楊純慧（2014）。PE092002 抑制胃癌細胞株生長之機制探討〔碩士論文，臺北醫學大學〕。華藝線上圖書館。https://doi.org/10.6831/TMU.2014.2014.00118
- 溫偉伶（2006）。組蛋白去乙醯?抑制劑抑制肺癌細胞生長之機制探討〔碩士論文，國立臺灣師範大學〕。華藝線上圖書館。https://www.airitilibrary.com/Article/Detail?DocID=U0021-0712200716121428
- Hou, Y. C. (2017). 二甲雙胍對口腔癌化學預防的效果—以口腔黏膜下纖維化症患者為例 [master's thesis, Kaohsiung Medical University]. Airiti Library. https://www.airitilibrary.com/Article/Detail?DocID=U0011-2808201717301000
- 吳宋恩（2013）。二苯甲醯基甲烷及其類似物抑制人類子宮頸癌細胞的轉移作用〔碩士論文，朝陽科技大學〕。華藝線上圖書館。https://www.airitilibrary.com/Article/Detail?DocID=U0078-2712201314043249
- Pan, K. L., Hsu, Y. C., Tung, C. W., Chang, S. T., Chung, C. M., & Lin, C. L. (2020). Inhibition of Notch Signaling Alleviated Diabetic Macrovasculopathy in an In Vitro Model. Acta Cardiologica Sinica, 36(5), 503-513. https://doi.org/10.6515/ACS.202009_36(5).20200210A