【作者】 方明；

【导师】 梅晓冬；

【作者基本信息】 安徽医科大学 ， 内科学， 2006， 硕士

【摘要】 目的：用阿霉素体外诱导的方法建立多药耐药细胞系A549／ADM。研究阿奇霉素对A549／ADM的耐药逆转作用。方法 采用阿霉素诱导A549使其耐药。首先在培养液中使用低浓度阿霉素持续诱导，再逐渐、间歇增加阿霉素浓度。用流式细胞仪（FCM）测定A549和A549／ADM的P-GP和MRP的表达。用MTT法分别检测四种药物对A549、A549／ADM、使用阿奇霉素（AZM）处理后的A549／ADM的半数抑制浓度(IC₅₀)。MTT法检测不同浓度阿奇霉素对A549／ADM的抑制率。用FCM检测A549、A549／ADM及使用阿奇霉素处理的A549／ADM细胞内ADM荧光强度。结果A549的P-GP和MRP的表达分别为2.3％和2.5％，而A549／ADM的P-GP和MRP的表达分别为33％和46.3％。A549／ADM对阿霉素的耐药指数为A549的17.80倍，对VCR、DDP、MTX均有不同程度的耐药。使用5μg／ml阿奇霉素处理后，A549／ADM对ADM的敏感性为原来的2.81倍，对DDP、VCR、MIX的敏感度分别为原来的1.86、3.78、3.86倍。加入10μg／ml的阿奇霉素后，A549／ADM对上述药物的敏感程度提高更明显。同样浓度的阿霉素作用相同时间后，A549、A549／ADM和使用阿奇霉素处理后的A549／ADM的荧光强度分别为121.4、51.2、96。结论 利用阿霉素在体外采用低浓度梯度持续诱导，加以高浓度间歇诱导的方法，可以建立多药耐药细胞系。耐药细胞系A549／ADM表现对多种常用化疗药物不同程度的耐药。P-GP、MRP的高表达在MDR的发生中可能起着重要作用。阿奇霉素可能通过增加药物在细胞内的聚集而逆转A549／ADM的多药耐药。其逆转效果与剂量相关。更多还原

【Abstract】 Objective To establish a multidrug-resistant cell line A549/ADM induced by azithromycin in vitro and investigate the reversal effect of azithromycin to A549/ADM. Methods A549 cells were treated with adriamycin （ADM）. The cells were treated with low concentration of ADM firstly and then gradually increased concentration of ADM intermittently . The expressions of P-GP and MRP in A549 and A549/ADM were confirmed by flow cytometry（FCM）. The 50% restrain concentration (IC₅₀) of four drugs to A549 and A549/ADM were evaluated with MTT assay. Same methods were used to A549/ADM when treated with azithromycin（AZM）. The inhibition rates of different concentration of AZM to A549/ADM were also evaluated with MTT assay. The fluorescence intensity of ADM in A549 and A549/ADM with or without AZM added were also evaluated with FCM. Results The expressions of P-GP and MRP in A549 were 2.3% and 2.5% respectively, but those of A549/ADM were 33% and 46.3% respectively.The resistance index of A549/ADM to ADM was 17.80 times higher than that of A549 and the cells showed resistant to DDP, VCR and MTX incoordinately .With AZM added（5μg/ml）, the sensitivities of A549/ADM to ADM were 2.81 times higher than before and sensitivities to DDP,VCR and MTX increased by 1.86, 3.78 and 3.86 times respectively. The sensitivities of A549/ADM to these drugs were much higher when treated with 10μg/ml of AZM. With the same treatment, the fluorescence intensities in A549 , A549/ADM and A549/ADM were 121.4, 51.2 and 96 respectively. Conclusions A549/ADM were multidrug- resistant（MDR） .The high expressions of P-GP and MRP might play an important role in MDR. AZM could reverse the MDR of A549/ADM through increase the accumulation of drugs in cells and the effection of reversal correlates with concentration.更多还原