Ferreira-Pinto K C, Miranda-Vilela A L, Anacleto C, Fernandes A P, Abdo M C, Petrillo-Peixoto M L, Moreira E S
Departamento de Microbiologia, Instituto de Ciências Biologicas, Universidade Federal de Minas Gerais, Brasil.
Can J Microbiol. 1996 Sep;42(9):944-9. doi: 10.1139/m96-121.
A glucantime sensitive Leishmania (V.) guyanensis strain was used to obtain in vitro resistant cell lines, by increments in glucantime concentrations employing both one step and stepwise protocols. Whereas the effective concentration of drug that inhibited the growth of wild type cells by 50% (EC50 value) was 0.20 mg Sb(v)/mL, the resistant cells were able to grow in glucantime concentrations greater than 8.0 mg/mL. The resistant cell lines were partially characterized by their in vitro response to glucantime, the stability of resistance phenotype, cross resistance to a range of drugs, and also by the analysis of total DNA fragments generated by restriction endonucleases and blot hybridization. Amplified DNA sequence similar to a P-glycoprotein analog from Leishmania tarentolae (ltpgpA gene) was observed in all the resistant cell lines obtained through the one-step protocol. These cell lines showed cross resistance to heavy metals but were sensitive to puromycin, vinblastine, and pentostam.
通过一步法和逐步递增法增加葡糖胺锑钠浓度，利用对葡糖胺锑钠敏感的圭亚那利什曼原虫（V.）菌株获得体外抗性细胞系。抑制野生型细胞生长50%的药物有效浓度（EC50值）为0.20mg Sb（v）/mL，而抗性细胞能够在浓度大于8.0mg/mL的葡糖胺锑钠中生长。通过对葡糖胺锑钠的体外反应、抗性表型的稳定性、对一系列药物的交叉抗性以及对限制性内切酶产生的总DNA片段的分析和印迹杂交，对抗性细胞系进行了部分表征。在通过一步法获得的所有抗性细胞系中，均观察到与来自热带利什曼原虫的P-糖蛋白类似物（ltpgpA基因）相似的扩增DNA序列。这些细胞系对重金属表现出交叉抗性，但对嘌呤霉素、长春碱和喷他脒敏感。
