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本研究通过建立人肝癌多药耐药细胞系,研究家蝇抗菌肽对该耐药细胞耐药性的逆转作用.采用阿霉素浓度梯度递增体外诱导法建立人肝癌耐药细胞株Bel-7402/ADM;采用MTT法检测阿霉素、顺铂、吉西他滨以及抗菌肽对细胞的毒性作用,并计算Bel-7402/ADM细胞对阿霉素、顺铂、吉西他滨耐药指数(RI);荧光显微镜观察细胞内阿霉素的荧光强度;采用流式细胞术检测细胞周期及细胞表面P-糖蛋白的含量.细胞毒性检测结果显示Bel-7402/ADM对阿霉素、顺铂和吉西他滨耐药指数分别为10.96、1.74和8.14,周期分析结果显示Bel-7402/ADM细胞的细胞周期较Bel-7402细胞发生改变:G0/G1期细胞比例明显升高,S期的细胞比例下降,荧光显微镜观察Bel-7402/ADM细胞内阿霉素量明显低于Bel-7402细胞,流式细胞术检测结果显示,Bel-7402/ADM细胞表面P-gp含量明显高于Bel-7402细胞,表明建成的Bel-7402/ADM细胞对多种药物耐药.抗菌肽对细胞的毒性作用检测显示,抗菌肽对Bel-7402/ADM细胞抑制作用呈剂量依赖性,但耐药细胞对抗菌肽并无耐药性.小剂量(100 mg/mL)的抗菌肽作用后,Bel-7402/ADM细胞的耐药指数减小至7.1,荧显微镜观察Bel-7402/ADM细胞内阿霉素荧光强度增强,流式细胞术检测Bel-7402/ADM细胞表面P-gp表达降低.结果表明,抗菌肽有可能通过降低细胞表面P-gp的表达起到逆转Bel-7402/ADM细胞耐药性的作用.“,”The aim of this study was to establish an adriamycin-induced multidrug resistance human hepatocellular carcinoma Bel-7402/ADM cells,and to investigate whether Musca domestica antimicrobial peptides could reverse the muhidrug resistance in vitro and insight the potential mechanisms involved.By increasing the concentration of adriamycin in vitro gradually,a multidrug resistance human hepatocellular carcinoma cell line Bel-7402/ADM was established.The resistance indexes (RI) of the cells were measured as high as 10.97 times by MTT assays compared with the Bel-7402 cells.Furthermore,Bel-7402/ADM cells were also shown resistance to cisplatin (1.74) and gemcitabine (8.14).Compared with the Bel-7402 cells,the percentages of cell in G0/G1 were significantly increased in Bel-7402/ADM,and those in S declined dramatically.The fluorescence intensity of intracellular adriamycin in Bel-7402/ADM cells was significantly lower than in Bel-7402 cells.P-gp of Bel-7402/ADM cells were higher than Bel-7402 cells under the same circumstance.After treatment with antimicrobial peptides,the resistance indexes (RI) of Bel-7402/ADM cells were declined to 7.1,the fluorescence intensity of intracellular adriamycin were increasing in Bel-7402/ADM cells,and the expression of P-gp was declined,which indicate that AMPs can reverse the multidrug resistance in Bel-7402/ADR cells by decline the expression of P-gp.