Uozurmi K, Nakaichi M, Yamamoto Y, Une S, Taura Y
Department of Veterinary Surgery, Yamaguchi University, School of Veterinary Science, 1677-1 Ooaza Yoshida, Yamaguchi 753-8515, Japan.
Res Vet Sci. 2005 Jun;78(3):217-24. doi: 10.1016/j.rvsc.2004.09.012. Epub 2005 Jan 26.
New multidrug resistant cell lines developed from the canine B cell lymphoma cell line (GL-1) were characterized in terms of chemosensitivity to some antineoplastics and P-glycoprotein (Pgp) expression. GL-1 was continuously exposed to a culture medium containing gradually increasing levels of doxorubicin and the cells that could grow in the presence of doxorubicin were obtained. Chemosensitivity of these cells to various antineoplastics were investigated with or without verapamil, which reversed Pgp-mediated drug resistance. The expression of Pgp on the cells was also examined by Western blot analysis. As a result, three kinds of resistant cell lines, designated as GL-DOX60, 300, and 4000 were obtained. These cell lines showed stable proliferation in the medium containing 60, 300, and 4000 ng/ml, respectively. These cells were much more resistant to vincristine than doxorubicin. This resistance was strongly reversed by the presence of verapamil. On the other hand, cisplatin was effective enough in killing these derived cells. In the Western Blot analysis, some bands that reacted to the anti-human Pgp monoclonal antibodies were observed in GL-DOX4000. The cells derived from GL-1 have multidrug resistance potential mediated by canine Pgp. The cells produced in this experimental trial are considered to be useful models for various investigations on canine multidrug resistance.
从犬B细胞淋巴瘤细胞系（GL-1）培育出的新型多药耐药细胞系，在对某些抗肿瘤药的化学敏感性和P-糖蛋白（Pgp）表达方面进行了特性鉴定。GL-1持续暴露于含有逐渐增加浓度阿霉素的培养基中，从而获得了能在阿霉素存在下生长的细胞。在有或没有维拉帕米（维拉帕米可逆转Pgp介导的耐药性）的情况下，研究了这些细胞对各种抗肿瘤药的化学敏感性。还通过蛋白质免疫印迹分析检测了细胞上Pgp的表达。结果，获得了三种耐药细胞系，分别命名为GL-DOX60、300和4000。这些细胞系分别在含有60、300和4000 ng/ml的培养基中显示出稳定增殖。这些细胞对长春新碱的耐药性比对阿霉素更强。维拉帕米的存在可强烈逆转这种耐药性。另一方面，顺铂对杀死这些衍生细胞足够有效。在蛋白质免疫印迹分析中，在GL-DOX4000中观察到了一些与抗人Pgp单克隆抗体发生反应的条带。源自GL-1的细胞具有由犬Pgp介导的多药耐药潜能。在该实验中产生的细胞被认为是用于犬多药耐药性各种研究的有用模型。
