Trinh Son Xuan, Nguyen Huyen Thi Bich, Saimuang Kween, Prachayasittikul Virapong, Chan On Waraporn
Center for Research and Innovation, Faculty of Medical Technology, Mahidol University, Bangkok, Thailand. Email:
Asian Pac J Cancer Prev. 2017 Feb 1;18(2):473-477. doi: 10.22034/APJCP.2017.18.2.473.
Background: Metformin is an oral anti-diabetic agent that has been widely prescribed for treatment of type II diabetes. Anti-cancer properties of metformin have been revealed for numerous human malignancies including cholangiocarcinoma (CCA) with anti-proliferative effects in vitro. However, effects on CCA cell migration and invasion have not been fully investigated. The present study aimed to explore the inhibitory effects of metformin on motility, migration and invasion of the CCA cell line HuCCT1, and examine molecular mechanisms underlying metformin effects. Methods: HuCCT1 cells were exposed to increasing doses of metformin. Viability and growth of HuCCT1 cells were assessed by MTS and colony formation assays, respectively. Motility, migration and invasion of metformin-treated HuCCT1 cells were determined in vitro using wound healing, transwell migration and matrigel invasion assays. Expression of signaling molecules and epithelial-mesenchymal transition (EMT) markers was assessed by Western blotting. Results: It was observed that metformin significantly decreased HuCCT1 cell viability and colony formation. The agent also markedly reduced wound closure, migration and invasion of HuCCT1 cells. Furthermore, metformin exposure resulted in decreased STAT3 activation and down-regulation of anti-apoptotic protein Bcl-2 and Mcl-1 expression. In addition, it upregulated the expression of E-cadherin, while downregulating that of N-cadherin, Snail, and MMP-2. Conclusion: These results demonstrated inhibitory effects of metformin on CCA cell migration and invasion, possibly involving the STAT3 pathway and reversal of EMT markers expression. They further suggest that metformin may be useful for CCA management.
二甲双胍是一种口服抗糖尿病药物，已被广泛用于治疗II型糖尿病。二甲双胍对多种人类恶性肿瘤具有抗癌特性，包括胆管癌（CCA），在体外具有抗增殖作用。然而，其对CCA细胞迁移和侵袭的影响尚未得到充分研究。本研究旨在探讨二甲双胍对CCA细胞系HuCCT1的运动性、迁移和侵袭的抑制作用，并研究其作用的分子机制。方法：将HuCCT1细胞暴露于不同剂量的二甲双胍中。分别通过MTS和集落形成试验评估HuCCT1细胞的活力和生长。使用伤口愈合试验、Transwell迁移试验和基质胶侵袭试验在体外测定经二甲双胍处理的HuCCT1细胞的运动性、迁移和侵袭。通过蛋白质免疫印迹法评估信号分子和上皮-间质转化（EMT）标志物的表达。结果：观察到二甲双胍显著降低了HuCCT1细胞的活力和集落形成。该药物还显著减少了HuCCT1细胞的伤口闭合、迁移和侵袭。此外，二甲双胍处理导致STAT3激活降低以及抗凋亡蛋白Bcl-2和Mcl-1表达下调。此外，它上调了E-钙黏蛋白的表达，同时下调了N-钙黏蛋白、Snail和MMP-2的表达。结论：这些结果证明了二甲双胍对CCA细胞迁移和侵袭的抑制作用，可能涉及STAT3途径以及EMT标志物表达的逆转。它们进一步表明二甲双胍可能对CCA的治疗有用。
