人肝癌顺铂耐药细胞株HepG2的建立

Establishment of drug-resistant hepatocellular carcinoma cell line HepG2

目的体外建立顺铂(DDP)诱导的人肝癌耐药细胞株(HepG2/DDP),研究其生物学特性。方法以采用药物大剂量冲击结合低剂量持续诱导方法诱导HepG2细胞株,建立人肝癌顺铂耐药细胞株HepG2/DDP;MTT法检测顺铂对HepG2和HepG2/DDP的半数抑制浓度(IC50)和耐药系数(RI),绘制细胞生长曲线及计算细胞倍增时间,并用流式细胞仪(FCM)检测细胞周期。结果历时5个月成功建成HepG2/DDP细胞株,MTT法检测DDP对HepG2的IC50为1.35μg/ml,HepG2/DDP的IC50为23.35μg/ml,RI达17生长曲线表明其增殖速度明显低于亲本HepG2人肝癌细胞,且倍增时间增加;细胞周期分析发现相对于亲本HepG2细胞,G0/G1期细胞减少、S期与G2/M期细胞增多。结论成功建立稳定耐药细胞株HepG2/DDP。

Objective To establish a drug-resistant hepatocellular carcinoma cell line and observe its biological characters .Methods The drug-resistant cell line HepG2 was induced using a concentration gradient of cisplatin （DDP） with exposed to intermittently.Drug sensitivity of HepG2 and HepG2/DDP were detected by MTT assay.The resistance index of the resistant cell line to DDP was determined by the methyl thiazolyl tetrazoliumassay （ MTT ） .Cell growth curve was painted by cell counting assay and the doubling time was accounted.Cycle distribution were measured by flow cytometry .Results The DDP-resistant cell line HepG2/DDP was established after 5 months and indicated drug-resistance to DDP, the drug resistance index being 17 respectively.The growing rate of resistant cells was obviously slower than that of the parental cells .The distributing proportion of HepG 2/DDP cells in G0/G1 phase was less than that of the parental cells , whereas the percentages of cells in G 2/M and S phase were significantly increased in HepG 2/DDP in comparison with those in HepG2.Conclusions A reliable drug-resistant human hepatoma cell line HepG 2/DDP is successfully established.