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建立耐药肝癌细胞模型 7721/Adm并研究其生物学特性。方法 :应用人肝癌细胞株 7721(简称7721细胞株 ) ,采用阿霉素 (Adm)高浓度间歇诱导法 ,建立耐药人肝癌细胞模型 7721/Adm(简称7721/Adm细胞株 )。观察该细胞的生长规律 ;用MTT法检测该耐药细胞模型的多药耐药性 ;以流式细胞术检测该耐药模型细胞周期中的分布、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)的表达等。结果 :1 经MTT法检测7721/Adm细胞较HCC 7721细胞的阿霉素半数抑制浓度 (IC50)增大99 9倍 ,并对多种抗癌药物产生耐药性。2 耐药细胞的倍增时间明显延长 ,S期细胞减少 ,G1、G2 期细胞增多。3 该耐药模型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)有非常显著的升高 (P<0 01) ,P gp 表达为94 6 %、MRP表达为69 4% ,而谷胱甘肽硫转移系统 (GSH/GST)的表达无明显变化 (P>0 05)。结论 :7721/Adm细胞是一个明确的多药耐药模型 ;具有耐药细胞的基本生物学特性 :同亲本细胞相比 ,IC50提高99 9倍、倍增时间延长4 2倍、细胞内药物蓄积明显降低、细胞周期分布发生变化 ;其多药耐药性与P gp、MRP的高表达有密切关系
A drug resistant hepatoma cell model 7721/Adm was established and its biological characteristics were studied. METHODS: Human hepatocellular carcinoma cell line 7721 (referred to as 7721 cell line) was used. Adriamycin (Adm) high concentration intermittent induction method was used to establish a resistant human hepatoma cell model 7721/Adm (referred to as 7721/Adm cell line). The growth pattern of the cells was observed; the multidrug resistance of the drug-resistant cell model was detected by MTT assay; the distribution in the cell cycle of the drug-resistant model and the multidrug resistance gene (mdr) on the cell surface were detected by flow cytometry. Expression product P glycoprotein (P gp), multidrug resistance-associated protein (MRP), and glutathione sulfur transfer system (GSH/GST) expression. RESULTS : 1 The half-maximal inhibitory concentration (IC50) of 7721/Adm cells compared with HCC 7721 cells was 99 9-fold increased by MTT assay, and resistant to various anti-cancer drugs. 2 The doubling time of drug-resistant cells was significantly prolonged, S-phase cells decreased, and G1 and G2 cells increased. 3 The expression products of multidrug resistance gene (mdr) on the surface of the resistant model cell P glycoprotein (P gp) and multidrug resistance-associated protein (MRP) were significantly increased (P<0 01), P gp The expression was 94.6% and the expression of MRP was 69.4%. There was no significant change in the expression of GSH/GST (P>0.05). Conclusion: 7721/Adm cells are a definitive multidrug resistance model; the basic biological characteristics of drug-resistant cells are: compared with the parental cells, the IC50 is 99 9-fold longer, the doubling time is prolonged 42-fold, and the intracellular drug accumulation is obvious. Decreased, cell cycle distribution changes; its multidrug resistance is closely related to high expression of P gp and MRP