Guarnaccia Laura, Navone Stefania E, Masseroli Matteo M, Balsamo Melissa, Caroli Manuela, Valtorta Silvia, Moresco Rosa M, Campanella Rolando, Schisano Luigi, Fiore Giorgio, Galiano Valentina, Garzia Emanuele, Appiani Giuseppe C, Locatelli Marco, Riboni Laura, Marfia Giovanni
Laboratory of Experimental Neurosurgery and Cell Therapy, Neurosurgery Unit, Foundation IRCCS Ca' Granda Ospedale Maggiore Policlinico, 20122 Milan, Italy.
Department of Clinical Sciences and Community Health, University of Milan, 20122 Milan, Italy.
Cancers (Basel). 2022 Mar 10;14(6):1412. doi: 10.3390/cancers14061412.
Glioblastoma is the most aggressive primary brain malignancy in adults, with a poor prognosis of about 14 months. Recent evidence ascribed to metformin (MET), an antihyperglycemic drug, the potential to reduce cancer incidence and progression, but the molecular mechanisms underlying these effects need to be better investigated.
Here, we tested the efficacy of MET on = 10 primary glioblastoma endothelial cells (GECs), by viability and proliferation tests, as MTT and Live/Dead assays, apoptosis tests, as annexin V assay and caspase 3/7 activity, functional tests as tube-like structure formation and migration assay and by mRNA and protein expression performed by quantitative real-time PCR analysis (qRT-PCR) and Western Blot, respectively.
Data resulting revealed a time- and μ-dependent ability of MET to decrease cell viability and proliferation, increasing pro-apoptotic mechanisms mediated by caspases 3/7. Also, MET impacted GEC functionality with a significant decrease of angiogenesis and invasiveness potential. Mechanistically, MET was able to interfere with sphingolipid metabolism, weakening the oncopromoter signaling promoted by sphingosine-1-phosphate (S1P) and shifting the balance toward the production of the pro-apoptotic ceramide.
These observations ascribed to MET the potential to serve as add-on therapy against glioblastoma, suggesting a repurposing of an old, totally safe and tolerable drug for novel oncology therapeutics.
胶质母细胞瘤是成人中最具侵袭性的原发性脑恶性肿瘤，预后较差，约为14个月。最近有证据表明，抗高血糖药物二甲双胍（MET）具有降低癌症发病率和进展的潜力，但这些作用背后的分子机制尚需进一步研究。
在此，我们通过MTT和活/死检测等活力和增殖试验、膜联蛋白V检测和半胱天冬酶3/7活性等凋亡试验、管状结构形成和迁移试验等功能试验，以及分别通过定量实时PCR分析（qRT-PCR）和蛋白质印迹法进行的mRNA和蛋白质表达，测试了MET对10种原发性胶质母细胞瘤内皮细胞（GEC）的疗效。
所得数据显示，MET具有时间和剂量依赖性降低细胞活力和增殖的能力，增加了由半胱天冬酶3/7介导的促凋亡机制。此外，MET影响GEC功能，显著降低血管生成和侵袭潜力。从机制上讲，MET能够干扰鞘脂代谢，削弱由1-磷酸鞘氨醇（S1P）促进的肿瘤促进信号传导，并使平衡转向促凋亡神经酰胺的产生。
这些观察结果表明MET有潜力作为胶质母细胞瘤的辅助治疗药物，提示将一种古老、完全安全且耐受性良好的药物重新用于新型肿瘤治疗。
