摘要
目的建立人卵巢癌拓扑替康(TPT)耐药细胞株(SKOV3/TPT),研究其生物学特性和耐药特点。方法模拟临床用药特点,采用大剂量TPT(2160ng/ml)冲击建成人卵巢癌SKOV3/TPT耐药细胞株。MTT法检测多药耐药性,流式细胞仪测定细胞周期、胞内药物的积聚及P糖蛋白(Pgp)表达,从生长曲线、光镜下形态、超微结构等方面比较耐药细胞和SKOV3细胞某些生物学特性的差异。结果成功建立了SKOV3/TPT耐药株,耐药指数为10·67,对喜树碱、米托蒽醌及长春新碱有明显的交叉耐药,对紫杉醇、顺铂、5-氟尿嘧啶、足叶乙甙和甲氨蝶呤敏感。与SKOV3细胞相比,SKOV3/TPT耐药细胞倍增时间延长;G2+M期的比例明显增加(P<0·01);细胞内TPT浓度明显减少(P<0·05);无Pgp过表达。结论SKOV3/TPT细胞株呈中等水平耐药,耐药性稳定,呈非典型的多药耐药特征;对TPT耐药的主要原因可能与细胞内药物浓度降低和细胞周期改变及凋亡有关,与Pgp的过表达无关。
Objective To establish the topotecan（TPT）- resistant cell line of human oophoroma （SKOV3/TPT）, and to investigate its biological features. Methods To simulate the condition in clinical chemotherapy, drug resistant SKOV3/TPT cell line of human oophoroma was established by challenging SKOV3 with a large dose of TPT （2160ng/ml）. Drug sensitivity was assessed by MTT assay. Light microscopy, electronic microscopy, trypan blue staining rejection, and cell counting were employed to determine the biological features of the cell line. Flow cytometry （FCM） was used to study the cell cycle distribution, the accumulation of intracellular drug, and the expression of P glucoprotein （Pgp）. Results SKOV3/TPT cells were established successfully, and the resistance index （RI） was 10. 67. They showed an obvious cross-resistance to CPT-11, MX and VCR, but still sensitive to taxol, cis-platin, VP16, MTX and 5-Fluorouracil. Comparing with the parent cells, their morphology was unchanged, but multiplication time was prolonged. The apoptosis rate and the proportion of G2＋M phase cells of resistant cells were significantly higher than that of their parent cells （P〈0. 01）. The concentration of intracellular TPT was obviously lower （P〈0.05）, but no over expression of Pgp was found. Conclusions SKOV3/TPT cell line is of an atypical phenotype with multi-drug resistance. The resistance of the cell line may be due to the lowered concentration of intracellular drugs, the change in cell cycle and apoptosis, but it may not be related to over expression of Pgp.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2005年第8期715-717,共3页
Medical Journal of Chinese People's Liberation Army
基金
首都医学发展科研基金(重点学科)资助课题(首都ZD199915)
关键词
卵巢肿瘤
拓扑替康
抗药性
ovarian neoplasms
topotecan
drug resistance
