移植人类恶性肿瘤细胞的线粒体作为在实验动物肺部形成肿瘤生长的方法

O. I. Kit, E. M. Frantsiyant, A. Shikhlyarova, I. V. Neskubina
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引用次数: 0

摘要

研究目的通过移植从人类恶性肿瘤中分离出的线粒体,在 Balb/c 裸鼠体内获得肺癌肿瘤。在高速冷冻离心机上使用差速离心法分离人类肺癌线粒体。线粒体样本用 0.9 % 的氯化钠溶液稀释至每只动物 3 毫克的蛋白质浓度。将新鲜分离的肺癌线粒体移植到雄性 Balb/c 裸小鼠(n = 6)的尾静脉中,每只小鼠在 0.4 毫升生理盐水中吸收 3 毫克蛋白质。雄性 Balb/c 裸系小鼠(n = 4 只)作为对照组,每天尾静脉注射一次 0.4 毫升生理盐水。所有动物在静脉移植从肺癌中分离的线粒体 4 个月后被断头台处死。将线粒体导入小鼠尾静脉后,动物的行为和覆盖物的颜色均无变化。没有发现呼吸系统问题,也没有发现引入外来蛋白质后的过敏反应。移植线粒体的小鼠在第 113 天至 123 天出现呼吸问题,体重下降≥25%。从一名非角化性鳞状细胞癌患者的肺部静脉移植线粒体后,在一只 Balb/c Nude 雄性小鼠的肺部制备物上,可以清楚地看到肿瘤结节均匀地形成,肺部空间充血活跃。患者和直接接受肿瘤线粒体的 Balb/c 裸鼠的肺实质中肿瘤生长的特征相似,都是由胞质发育不良的鳞状上皮非典型多形性细胞的领域、细胞和紧密贴合的股组成的实体结构。此外,腺癌的迹象还包括腺状和乳头状结构以及带有大核基的细胞复合体。鳞状上皮细胞具有严重的不典型性和多形性。大多数细胞核形状不规则,核仁较大,颜色不均,核仁增大。多核细胞和病理有丝分裂的频率相当高,其分裂图形清晰可见,证明细胞具有增殖活性。将从恶性人类肺肿瘤中分离出的线粒体静脉移植到雄性 Balb/c 裸系小鼠体内,会导致雄性 Balb/c 裸系小鼠体内恶性病灶的生长和发展。这是以前在实验肿瘤学中不为人知的事实,从而可以研究人类癌细胞线粒体的新特性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Transplantation of mitochondria from human malignant tumor cells as a method of tumor growth formation in the lungs of laboratory animals
Purpose of the study. Obtaining lung cancer tumors in Balb/c Nude mice by transplantation of mitochondria isolated from human malignant tumors.Materials and methods. Human lung cancer mitochondria were isolated using differential centrifugation on a high-speed refrigerated centrifuge. Mitochondrial samples were diluted with 0.9 % NaCl solution to a protein concentration of 3 mg per animal. Freshly isolated lung cancer mitochondria were once transplanted into the tail vein of male Balb/c Nude mice (n = 6) at the rate of 3 mg of protein per 1 animal in 0.4 ml of saline. Male mice of Balb/c Nude line (n = 4), which were injected with 0.4 ml of physiological solution into the tail vein once a day, served as a control group. All animals were killed by guillotine decapitation 4 months after intravenous transplantation of mitochondria isolated from lung cancer.Results. After the introduction of mitochondria into the tail vein of mice, no changes were found in the animals’ behavior, coloration of coverings. No respiratory problems, and no allergic reactions to the introduction of foreign protein were detected. Respiratory problems in mice with mitochondrial transplantation occurred between days 113 and 123, when a drop in body weight ≥ 25 % was observed. A uniform formation of tumor nodes together with an active filling of the lung space with blood was clearly visible on lung preparations of a Balb/c Nude male mouse after intravenous transplantation of mitochondria from the lung of a patient with non-keratinizing squamous cell carcinoma. Similar characteristics of tumor growth in the lung parenchyma of the patient and Balb/c Nude mice as direct recipients of tumor mitochondria involved solid structures consisting of fields, cells and strands of tightly fitting atypical polymorphic cells of the squamous epithelium with poorly developed cytoplasm. In addition, signs of adenocarcinoma included glandular and papillary-like structures and cell complexes with large nucleoli. Squamous epithelial cells were characterized by severe atypia and polymorphism. The shape of the nuclei was irregular in most cells, with large sizes, the color was heterogeneous, and the nucleoli were enlarged. Multinucleated cells, as well as pathological mitoses with clearly marked division figures, which testified to their proliferative activity, were encountered with a rather high frequency.Conclusions. Application of intravenous transplantation of mitochondria isolated from malignant human lung tumor in male mice of Balb/c Nude line causes growth and development of malignant foci in male Balb/c Nude mice. This is previously unknown fact in experimental oncology, allowing the study of new properties of mitochondria of human cancer cells.
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