人肝细胞癌耐阿霉素细胞亚株SMMC-7721/ADM的诱导及其特征研究

Establishment of Adriamycin-Resistance Cell Substrain SMMC-7721/Adriamycin of Human Hepatocellular Carcinoma and Research of Its Characteristics

目的 建立人肝细胞癌多药耐药亚株并对其主要特征进行研究。方法 应用SMMC 772 1细胞株 ,通过不断提高培养液中阿霉素 (ADM)的浓度 ,长期筛选培养 ,得到了人肝细胞癌多药耐药亚株SMMC 772 1/ADM后 ,对其特征进行研究。结果 ①SMMC 772 1/ADM对ADM的抗药性提高了 33.3倍 ,对长春新碱的抗药性提高了 16 .8倍 ,对顺铂的抗药性提高了 2 .8倍 ;②耐药细胞与亲代细胞的生长速度基本相同 ,倍增时间分别为 32 .0h和 30 .5h ,呈现相似的生长曲线 ;③利用扫描和透射电镜发现 ,耐药细胞与亲代细胞形态上的明显差别是前者微绒毛变得粗短、稀疏、分布不均 ;④耐药细胞保持着肝细胞癌的特征 ,能在裸小鼠皮下移植再现 ;⑤SMMC 772 1/ADM在去药培养 1个月和 2个月后耐药指数下降为 2 8.0 %和 9.2 % ,而在ADM( 0 .0 4 μg/ml)存在下生长 2个月后 ,耐药倍数可保持稳定 ( 35 .4倍 )。结论 SMMC 772 1/ADM细胞亚株具有稳定的耐药性细胞株的基本特征。

Objective To establish multidrug resistance cell substrain of human hepatocellular carcinoma and to investigate its characteristics.Methods SMMC 7721 cell strain was cultured in Adriamycin(ADM). The multidrug resistance cell substrain SMMC 7721/ADM was harvested after a long period of culture by gradually increasing the concentration of ADM and its characteristics were investigated. Results ①The drug resistance of SMMC 7721/ADM to ADM increased by 33.3 times, to Vincristine 16.8 times, to Diamminedichloroplatinum 2.8 times. ②The drug resistance cell substrain had almost the same growth velocity as its parental generation. The doubling time was 32.0 hours and 30.5 hours respectively. They had the analogous growth curves. ③The obvious difference between the drug resistance cell substrain and its parental generation was that the former’s microvilli became thick, short and scattered by scanning and transmitting electron microscopy. ④The multidrug resistance cell substrain kept the characteristics of hepatocellular carcinoma, it could be transplanted into the subcutaneous tissue of nude mice. ⑤The drug resistance of the cell substrain reduced to 28.0% and 9.2%after removal of the drug for 1 month and 2 months respectively, its drug resistance could remain stable (35.4 times) after 2 months of culture in ADM (0.04 μg/ml).Conclusion The SMMC 7721/ADM cell substrain has the stable fundamental characteristics of a drug resistance cell strain.