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目的 在体内建立人肝癌裸小鼠原位移植多药耐药模型 ,并研究其耐药机理。方法采用人肝癌 (BEL 740 2 )裸小鼠 (4～ 6周龄 )原位移植 ,给予阿霉素 (1.5mg/kg体重 ,每周 1次 )腹腔注射诱导耐药 (共 8周 ) ,经四唑蓝 (MTT)快速比色法检测原代培养的耐药细胞对抗癌药的敏感性 ,以流式细胞仪检测癌细胞表面mdr1基因产物P 糖蛋白 (P gp)的表达 ,并用罗丹明试验观察该蛋白功能。结果 移植瘤组织形态及生物学方面符合人肝癌特征 ,实验组肝癌细胞表面P gp表达为 (75 .45± 5 .67) % ,而对照组表达仅 (4.2 5± 1.2 8) % ,差异有非常显著性 (P <0 .0 1) ,对阿霉素的耐药倍数提高了16.67倍 ,对羟基喜树碱具有交叉耐受性 (13 .67倍 )。耐药细胞表面P gp有较强的药物外排功能。结论 阿霉素较易诱导原位移植于裸小鼠的人肝癌多药耐药性的产生 ,其耐药倍数与临床肝细胞癌相似。该模型的建立对研究肝癌多药耐药的产生机制以及逆转其多药耐药性具有重要价值
Objective To establish a multidrug resistance model for orthotopic transplantation of human hepatocellular carcinoma nude mice in vivo and study its drug resistance mechanism. Methods Human liver cancer (BEL 740 2 ) nude mice (4-6 weeks old) were orthotopically transplanted, and doxorubicin (1.5 mg/kg body weight, once weekly) was induced by intraperitoneal injection to induce drug resistance (8 weeks). The sensitivity of primary cultured drug-resistant cells to anti-cancer drugs was detected by MTT colorimetric assay. The expression of mdr1 gene product P glycoprotein (P gp) was detected by flow cytometry. Rhodamine test to observe the function of the protein. Results The morphology and biology of the transplanted tumors were consistent with the features of human liver cancer. The expression of P gp in the experimental group was (75.45 ± 5.67) % compared with that in the control group (4.2 5 ± 1.2 8) %. Significantly (P < 0.01), the resistance to adriamycin was increased by 16.67-fold and cross-tolerance to hydroxycamptothecin (13.67-fold). Pgp on the surface of drug-resistant cells has a strong drug efflux function. Conclusion Doxorubicin is more likely to induce the development of multidrug resistance in human hepatocellular carcinoma after orthotopic transplantation in nude mice. The multiple drug resistance is similar to that of clinical hepatocellular carcinoma. The establishment of this model is of great value for studying the mechanism of multidrug resistance in hepatocellular carcinoma and reversing its multidrug resistance.