Zhong Guo-xing, Gong Yi, Yu Chuan-jiang, Wu Shi-fei, Ma Qing-ping, Wang Yu, Ren Jiang, Zhang Xue-chao, Yang Wei-han, Zhu Wen
State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, 610041, China.
Tumour Biol. 2015 Jun;36(6):4689-97. doi: 10.1007/s13277-015-3117-8. Epub 2015 Jan 27.
Low molecular weight heparin (LMWH) improving the cancer survival has been attracting attention for many years. Our previous study found that LMWH (Fraxiparine) strongly downregulated the invasive, migratory, and adhesive ability of human lung adenocarcinoma A549 cells. Here, we aimed to further identify the antitumor effects and possible mechanisms of Fraxiparine on A549 cells and human highly metastatic lung cancer 95D cells. The ability of cell invasion, migration, and adhesion were measured by Transwell, Millicell, and MTT assays. FITC-labeled phalloidin was used to detect F-actin bundles in cells. Chemotactic migration was analyzed in a modified Transwell assay. Measurement of protein expression and phosphorylation activity of PI3K, Akt, and mTOR was performed with Western blot. Our studies found that Fraxiparine significantly inhibited the invasive, migratory, and adhesive characteristics of A549 and 95D cells after 24 h incubation and showed a dose-dependent manner. Fraxiparine influenced the actin cytoskeleton rearrangement of A549 and 95D cells by preventing F-actin polymerization. Moreover, Fraxiparine could significantly inhibit CXCL12-mediated chemotactic migration of A549 and 95D cells in a concentration-dependent manner. Furthermore, Fraxiparine might destroy the interaction between CXCL12-CXCR4 axis, then suppress the PI3K-Akt-mTOR signaling pathway in lung cancer cells. For the first time, our data indicated that Fraxiparine could significantly inhibit the motility of lung cancer cells by restraining the actin cytoskeleton reorganization, and its related mechanism might be through inhibiting PI3K-Akt-mTOR signaling pathway mediated by CXCL12-CXCR4 axis. Therefore, Fraxiparine would be a potential drug for lung cancer metastasis therapy.
低分子量肝素（LMWH）改善癌症生存率多年来一直备受关注。我们之前的研究发现，LMWH（速碧林）强烈下调人肺腺癌A549细胞的侵袭、迁移和黏附能力。在此，我们旨在进一步确定速碧林对A549细胞和人高转移性肺癌95D细胞的抗肿瘤作用及可能机制。通过Transwell、Millicell和MTT试验检测细胞侵袭、迁移和黏附能力。用异硫氰酸荧光素（FITC）标记的鬼笔环肽检测细胞中的F-肌动蛋白束。在改良的Transwell试验中分析趋化性迁移。用蛋白质印迹法检测PI3K、Akt和mTOR的蛋白表达及磷酸化活性。我们的研究发现，孵育24小时后，速碧林显著抑制A549和95D细胞的侵袭、迁移和黏附特性，并呈剂量依赖性。速碧林通过阻止F-肌动蛋白聚合影响A549和95D细胞的肌动蛋白细胞骨架重排。此外，速碧林能以浓度依赖性方式显著抑制CXCL12介导的A549和95D细胞趋化性迁移。此外，速碧林可能破坏CXCL12-CXCR4轴之间的相互作用，进而抑制肺癌细胞中的PI3K-Akt-mTOR信号通路。我们的数据首次表明，速碧林可通过抑制肌动蛋白细胞骨架重组显著抑制肺癌细胞的运动性，其相关机制可能是通过抑制CXCL12-CXCR4轴介导的PI3K-Akt-mTOR信号通路。因此，速碧林将是一种潜在的肺癌转移治疗药物。
