PFC体内递增诱导建立S180肿瘤多药耐药模型及其稳定性观察  

Establishment of S180 Tumor Multidrug Resistance Mouse Model by Increasing PFC and Observation on Stability

作　　者：顾云浩 曹晨洁 胡碧原 王俊[1] 韩东冬[1] 许爱华[1] 

机构地区：[1]扬州大学医学院,扬州225001

出　　处：《实验动物与比较医学》2015年第5期367-373,共7页Laboratory Animal and Comparative Medicine

基　　金：扬州大学大学生科技创新基金项目（2014）;江苏省医药高技术研究项目（BG2007609）

摘　　要：目的小鼠体内诱导建立获得性S180多药耐药（multi—drug resistance，MDR）模型及其稳定性观察。方法模拟临床顺铂＋氟尿嘧啶＋环磷酰胺（PFC）化疗方案给药，分三个阶段剂量递增法诱导S180腹水瘤小鼠，建立获得性S180MDR实验模型。采用噻唑蓝fMTTl法、流式细胞术动态检测各阶段所诱导细胞对化疗药物的耐药倍数、细胞内药物积累量及细胞膜P-糖蛋白（P-glycoprotein，P—gp）功能活性，并通过检测以上指标观察各阶段所诱导细胞停药后的耐药稳定性：采用实时荧光定量PCR（RT—qPCR）法检测各阶段所诱导细胞MDR-1 mRNA、多药耐药相关蛋白-1（multidrug resistance-associated proteinl，MRP-1）mRNA的表达量。结果与亲本细胞对照组比较，各阶段所诱导S180细胞对化疗药物的耐药倍数随着诱导时间延长和剂量增高而逐渐增大，细胞内阿霉素（adriamycin，ADR）积累量逐渐减少，细胞P—gp功能活性逐渐增强;各阶段所诱导S180细胞MDR-1mRNA、MRP-1 mRNA的表达量也与诱导时间和给药剂量呈正相关：第一、二和三阶段所诱导细胞的稳定耐药时间分别为1周、2周和3周左右。结论模拟临床PFC化疗方案给药，采用分阶段剂量递增小鼠体内诱导法可建立耐药强度高、稳定时间长的获得性S180MDR实验模型。Objective To establish aquired S 180 multidrug resistance （MDR） mouse model and study on its stability. Methods To mimic the clinical PFC （cis-Dichlorodiamineplatinum＋5- Fluorouracil＋cyclophosphamide） scheme, gradually increase the dose in three phases to induce S 180 ascites tumor mice and establish the aquired S 180 MDR mouse model. The induced cells resistance factor to the chemotherapeutics drugs in different stages, the accumulation of adriamycin （ADR） and the functional activity of P-glycoprotein （P-gp） were detected by MTT assay and flow cytometry. And then through detecting the above indicators to monitor the resistance drug stability of induced cells in different stages. The mRNA expression of MDR-1 and multidrug resistance-associated protein 1 （MRP-1） of induced cells in different stages were detected by real time quantitative PCR （RT-qPCR）. Results Compared with the parent cells, with induction time extending and dose increasing, the resistance factors in each stage of induced S 180 ceils to chemotherapeutics drugs were gradually increased, the accumulation of ADR was gradually reduced, and the functional activity of P-gp was strengthened. The mRNA expression of MDR-1 and MRP-1 of induced cells in different stages had a positive correlation to the induction time and dose. The stable resistance time of induced cells in the first, second and third phase are respectively for about 1 week, 2 weeks and 3 weeks. Conclusion To mimic the clinical PFC scheme, using the dose gradually increasing by phased can establish a high resistant strength, long stable time aquired S 180MDR experimental model.

关 键 词：顺铂+氟尿嘧啶+环磷酰胺(PFC) 多药耐药(MDR) S180细胞株 体内 细胞膜P-糖蛋白(P-gp) MDR-1 MRNA 多药耐药相关蛋白-1(MRP-1) MRNA 

分 类 号：R73-3[医药卫生—肿瘤]