Agnetti Lucrecia, Fondello Chiara, Villaverde Marcela S, Glikin Gerardo C, Finocchiaro Liliana M E
Unidad de Transferencia Genética Instituto de Oncología "Ángel H. Roffo" Universidad de Buenos Aires, Buenos Aires, Argentina.
Oncoscience. 2017 Dec 28;4(11-12):199-214. doi: 10.18632/oncoscience.387. eCollection 2017 Nov.
We originated and characterized melanoma cell lines derived from tumors of two feline and two canine veterinary patients. These lines reestablished the morphology, physiology and cell heterogeneity of their respective parental tumors. We evaluated the cytotoxicity of bleomycin (BLM) alone, or combined with interferon-β (IFN-β) or HSVtk/GCV suicide gene (SG) lipofection on these cells. Although the four animals presented stage III disease (WHO system), SG treated feline tumors displayed stable disease in vivo, while the canine ones exhibited partial response. Their derived cell lines reflected this behavior. Feline were significantly more sensitive than canine cells to IFN-β gene transfer. BLM improved the antitumor effects of both genes. The higher levels of reactive oxygen species (ROS) significantly correlated with membrane and DNA damages, emphasizing ROS intervention in apoptotic and necrotic cell death. After 3 days of BLM alone or combined with gene treatments, the colony forming capacity of two canine and one feline treatments survivor cells almost disappeared. Taken together, these results suggest that the treatments eradicated tumor initiating cells and support the clinical potential of the tested combinations.
我们从两只猫科和两只犬科兽医患者的肿瘤中分离并鉴定了黑色素瘤细胞系。这些细胞系重现了其各自亲本肿瘤的形态、生理和细胞异质性。我们评估了单独使用博来霉素（BLM），或与干扰素-β（IFN-β）或单纯疱疹病毒胸苷激酶/丙氧鸟苷自杀基因（SG）脂质转染联合使用时对这些细胞的细胞毒性。尽管这四只动物均表现为III期疾病（世界卫生组织系统），但接受SG治疗的猫科肿瘤在体内显示病情稳定，而犬科肿瘤则表现出部分缓解。它们衍生的细胞系反映了这种行为。猫科细胞比犬科细胞对IFN-β基因转移明显更敏感。BLM增强了两种基因的抗肿瘤作用。较高水平的活性氧（ROS）与膜和DNA损伤显著相关，强调了ROS在凋亡和坏死性细胞死亡中的干预作用。单独使用BLM或与基因治疗联合使用3天后，两只犬科和一只猫科治疗存活细胞的集落形成能力几乎消失。综上所述，这些结果表明这些治疗方法根除了肿瘤起始细胞，并支持所测试联合治疗方法的临床潜力。
