耐药人肝癌细胞模型-7721/Adm的建立

Establishment of human multidrug-resistant hepatocellular carcinoma cell line(7721/Adm)

目的 培养建立耐药肝癌细胞模型 - 772 1/ Adm并研究其生物学特性 .方法 应用人肝癌细胞株 - 772 1(以下简称772 1细胞 ) ,采用阿霉素 (Adm )大剂量间歇诱导法 ,建立耐药人肝癌细胞模型 - 772 1/ Adm (以下简称 772 1/ Adm ) .观察该细胞的生长规律 ;用 MTT法鉴定该耐药细胞模型对多种抗癌药物的多药耐药性 ;以流式细胞技术检测该耐药细胞模型细胞周期分布、细胞表面多药耐药基因 (MDR)的表达产物 - P-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)的表达等 .结果 1经 MTT法鉴定 772 1/ Adm细胞较 HCC- 772 1细胞的阿霉素半数致死浓度 (IC5 0 )增大4.6 5倍 ,并对多种抗癌药物产生耐药性 ,其耐药性提高了 2～6倍不等 ;2耐药细胞倍增时间明显延长 ,S期细胞减少 ,而G2期细胞增多 ;3该耐药模型 P- gp,MRP表达有非常显著的升高 (P<0 .0 1) :P- gp表达为 94.6 % ,MRP表达为 6 9.4% ,而 GSH/ GST的表达无明显变化 (P<0 .0 5 ) .结论 1HCC-772 1/ Adm是一个明确的多药耐药细胞模型 ;2 772 1/

AIM To establish human multidrug resistant hepatocellular carcinoma cell line (7721/Adm) and to study its characteristics. METHODS Using intermittent administration of high dose ADM, human hepatocellular carcinoma cell line (7721) was induced to human multidrug resistant hepatocellular carcinoma cell line (7721/Adm). The multidrug resistance of 7721/Adm to multianticarcinogen was detected by MTT assay, and the distribution of its cell cycle, the expressions of P gp, multidrug resistance associated protein (MRP) and GSH/GST were also detected by flow cytometry. RESULTS ①7721/Adm was resistant to many anti tumor agents. Its IC50 of ADM was 4.65 times higher than that of 7721 and its drug resistance to ADM was 2-6 times higher than that of 7721; ②The multiplication time of 7721/Adm prolonged, and the number of cells in S phase decreased while that in G1 and G2 phase increased; ③The expressions of P gp and MRP were enhanced significantly (94.6% and 69.4% respectively), but the expression of GSH/GST kept stable. CONCLUSION ①We successfully established a multidrug resistant cell line (7721/Adm) which has the basic characteristics of drug resistance cells; ②Compared with that of its parent cell line 7721, IC50 of 7721/Adm increased 4.65 times, and its multiplication time prolonged 4.6 times, drug accumulation decreased remarkably and distribution of its cell cycle changed.