人肝癌裸鼠移植模型阿霉素诱导法建立肝癌多药耐药细胞系

目的:建立人肝癌多药耐药细胞系研究其耐药特性及机制.方法:给予移植人肝癌细胞的裸小鼠腹腔注射阿霉素(ADM)长期诱导,获得多药耐药细胞系BEL-7402/ADM.相差显微镜和HE染色观察细胞,MTT法检测耐药细胞的多药耐药性,流式细胞术检测耐药细胞表面多药耐药基因(mdr)的表达产物P糖蛋白(P-gp)、多药耐药相关蛋白(MRP)及谷胱甘肽硫转移系统(GSH/GST)的表达.结果:BEL-7402/ADM对多种抗癌药物产生耐药,对阿霉素的耐药性提高了20.33倍.BEL-7402/ADM细胞表面P-gp和MRP表达阳性,与对照细胞BEL-7402表达比较,差异十分显著(P<0.01).GSH/GST表达无明显变化.结论:BEL-7402/ADM具有明确的多药耐药性,该模型的建立对研究肝癌多药耐药的产生及逆转具有重要价值.

Establishment of a multidrug resistance cell line of human hepatic carcinoma in a transplantation nude mice model treated with adriamycin

Objective: To establish a multidrug resistan ce cell line of human hepatic carcinoma in a nude mice model with subcutaneous B EL-7402 cell implantion treated with adriamycin and to study the mechanism and characteristics of multidrug resistance. Methods: Through the long-term induction with injection of adriamycin to nude mice, we obtained the multidrug resistance cell line, BEL-7402/ADM.The cells were observed with phese contrast microscope and HE staining. The multidrug resistance was detected by u sing MTT assay. The expression of Mdr-1 gene-coded P-glycoprotein (P-gp), mu ltidrug resistance-associated protein (MRP), and GSH/GST were examined by using flow cytometric assay. Results: BEL 7402/ADM was resistant to many anti-tumor agents. The IC50 of ADM was 20.33 times higher than that of BE L-7402. The P-gp and MRP expressed positively on the surface of BEL-7402/ADM cells. Compare with the control, BEL-7402 cells, the difference was significant . Conclusion: BEL-7402/ADM was a model with multidrug resista nce and was useful for the study of mechanism of mutidrug resistance and reversi ng mutidrug resistance in hepatic carcinoma.