顺铂体外诱导的人肝癌耐药细胞株(QGY/DDP)的建立  

Establishment of a drug resistant human hepatoma cell line (QGY/DDP) induced by cisplatin in vitro

作　　者：杨俊霞[1] 袁成福[2] 罗映[1] 邱红梅[1] 汤为学[3] 

机构地区：[1]重庆医科大学药理学教研室,重庆400016 [2]湖北民族学院医学院生物化学教研室 [3]重庆医科大学病理生理学教研室

出　　处：《解放军医学杂志》2007年第11期1145-1147,共3页Medical Journal of Chinese People's Liberation Army

摘　　要：目的建立顺铂(DDP)诱导的人肝癌耐药细胞株(QGY/DDP),研究其生物学特性和耐药机制。方法采用间歇诱导法,逐步递增DDP浓度,获得耐药细胞株QGY/DDP;MTT法测定药物敏感性;细胞计数法计算倍增时间,流式细胞术检测细胞周期;原子吸收光谱法测定细胞内铂离子蓄积量,流式细胞仪测定P-糖蛋白(P-gp)和谷胱甘肽S-转移酶-π(GST-π)的表达水平。结果成功建立了顺铂诱导的人肝癌耐药细胞株QGY/DDP,耐药指数为10.81,与5-氟尿嘧啶、长春新碱等抗癌药有明显的交叉耐药;与QGY细胞相比,QGY/DDP细胞增殖减慢、倍增时间延长,G0/G1期细胞增多、S期与G2/M期细胞减少,细胞内Pt含量降低,GST-π的表达升高,而P-gp无明显变化。结论QGY/DDP细胞具有耐药表型及耐药细胞的生物学特性;其耐药机制与细胞内Pt含量降低和GST-π过表达有关,而与P-gp无关。Objective To establish a drug resistant human hepatoma cell line （QGY/DDP） induced by cisplatin （DDP） in vitro, and to investigate its biological features and mechanisms of resistance. Methods The drug resistant cell line （QGY/DDP） of hepatoma was established by intermittent administration of stepwise increas of the dosage of DDP. Drug sensitivity was evaluated by MTT assay. Cell counting was employed to graph the growth curve and to calculate the doubling time. Flow cytometry （FCM） was used to study the cell cycle distribution. In addition, the intracellular platinum accumulation was detected by atomic absorption spectrometry, and the expressions of P-glycoprotein （P-gp） and glutathione Stransferasen （GST-π） were analyzed by FCM. Results QGY/DDP cell line was established successfully after 3 months with stable resistance to DDP, and the resistance index （RI） was 10. 81. The established cell line exhibited obvious crosresistance to 5-FU, VCR, MMC, EPI and HCPT. Compared with parental cell line, the QGY/DDP cell line grew slower and its doubling time became longer. The proportion of G0/G1 phase cells of the resistant cells was significantly higher than that of their parental cells, whereas the proportion of S and C2/M phase cells was decreased. The cellular platinum accumulation was obviously decreased in the QGY/DDP cell line, and the expression of GST-n in QGY/DDP cells was higher than that of their parental cells, but no over expres- sion of P-gp was found in the resistant cell line. Conclusions QGY/DDP cell line shows the typical and stable resistant phenotype and possesses the basic biological features of resistant cells. The resistance of the cell line to DDP may be due to the reduction of intracellular platinttm accttmulation and the over expression of GST-π, but may not be related to the expression of P-gp.

关 键 词：肿瘤细胞 培养的 顺铂 药物耐受性 细胞系 

分 类 号：R349.5[医药卫生—基础医学]