Department of Pharmacy, Faculty of Medicine, University of Novi Sad, 21000 Novi Sad, Serbia.
Department of Histology and Embryology, Faculty of Medicine, University of Novi Sad, 21000 Novi Sad, Serbia.
Biomed Pharmacother. 2020 Oct;130:110510. doi: 10.1016/j.biopha.2020.110510. Epub 2020 Jul 21.
We investigated the effect of nitroglycerin with metformin on fibrosarcoma in hamsters. Syrian golden hamsters of both sexes, weighing approximately 60 g, were randomly allocated to control and experimental groups, with 8 animals per group. In all groups, 2 × 10 BHK-21/C13 cells in 1 ml were injected subcutaneously into the animals' backs. Peroral treatment carried out with nitroglycerin 25 mg/kg daily, or with metformin 500 mg/kg daily, or with a combination of nitroglycerin 25 mg/kg and metformin 500 mg/kg daily. Later validation experiments were conducted with double doses of the single therapy and additional rescue doses of mebendazole 460 mg/kg daily, via a gastric probe after tumor inoculation. After 2 weeks, when the tumors were approximately 2-3 cm in the control group, all animals were sacrificed. Blood samples were collected for hematological and biochemical analyses, the tumors were excised and weighed, and their diameters and volumes were measured. The tumor samples were pathohistologically and immunohistochemically assessed for proliferation marker protein Ki-67, proliferating cell nuclear antigen PCNA, hematopoietic progenitor cell antigen CD34, cluster of differentiation 31 (CD31), cytochrome c oxidase subunit 4 (COX4), mitochondria marker Cytochrome C, glucose transporter 1 (GLUT1) and inducible nitric oxide synthase (iNOS), and the main organs were toxicologically tested. The Ki-67 and PCNA positivity and the cytoplasmic marker (CD34, CD31, COX4, Cytochrome C, GLUT1, iNOS) immunoexpression in the tumor samples were quantified. The combination of nitroglycerin and metformin significantly inhibited fibrosarcoma growth in hamsters without toxicity, compared to monotherapy or control. The results were validated and confirmed in the subsequently accomplished experiment with doubled doses of the single drug therapy and in the rescue experiment with addition of mebendazole. The single treatments did not show significant antisarcoma effect, regardless of the dose. Co-treatment with mebendazole inhibited anticancer activity of the nitroglycerin and metformin combination. Mebendazole rescued tumor progression suppressed by the combination of nitroglycerin and metformin. Administration of nitroglycerin with metformin might be an effective and safe approach in novel nontoxic adjuvant and relapse prevention anticancer treatment.
我们研究了硝酸甘油与二甲双胍对仓鼠纤维肉瘤的影响。将体重约 60g 的雄性和雌性叙利亚金黄仓鼠随机分配到对照组和实验组，每组 8 只动物。所有组的动物背部皮下注射 1ml 中含有 2×10 BHK-21/C13 细胞。经口给予硝酸甘油 25mg/kg 每日、二甲双胍 500mg/kg 每日或两者联合治疗，每日 1 次。在肿瘤接种后，通过胃探针给予双剂量单药治疗和额外的甲苯咪唑 460mg/kg 每日解救剂量，进行后续验证实验。在对照组中，当肿瘤约为 2-3cm 时，所有动物均被处死。采集血液样本进行血液学和生化学分析，切除肿瘤并称重，测量其直径和体积。对肿瘤样本进行组织病理学和免疫组织化学评估，检测增殖标志物蛋白 Ki-67、增殖细胞核抗原 PCNA、造血祖细胞抗原 CD34、分化簇 31（CD31）、细胞色素 c 氧化酶亚基 4（COX4）、线粒体标记物细胞色素 C、葡萄糖转运蛋白 1（GLUT1）和诱导型一氧化氮合酶（iNOS）。对主要器官进行毒理学检测。对肿瘤样本中的 Ki-67 和 PCNA 阳性率和细胞质标志物（CD34、CD31、COX4、细胞色素 C、GLUT1、iNOS）免疫表达进行定量分析。与单药治疗或对照组相比，硝酸甘油和二甲双胍联合显著抑制了仓鼠纤维肉瘤的生长，且无毒性。该结果在随后进行的加倍单药治疗实验和添加甲苯咪唑的解救实验中得到了验证和确认。单独使用任何一种药物治疗均未显示出显著的抗肉瘤作用，无论剂量如何。与甲苯咪唑联合使用会抑制硝酸甘油和二甲双胍联合的抗癌活性。甲苯咪唑解救了由硝酸甘油和二甲双胍联合抑制的肿瘤进展。联合使用硝酸甘油和二甲双胍可能是一种有效且安全的方法，可用于新型非毒性辅助和复发预防抗癌治疗。
