卵巢癌拓扑替康耐药细胞株的建立及其生物学特性

目的 :建立卵巢癌拓扑替康 (TPT)耐药细胞株 ,研究其生物学特性。方法 :用浓度梯度递增法建立卵巢癌TPT耐药细胞株。用四甲基偶氮唑蓝 (MTT)比色法检测此耐药株对多种化疗药物的耐药指数 ;用细胞计数法描绘两株细胞的生长曲线并计算群体倍增时间 ;用流式细胞仪检测两株细胞的凋亡率及周期分布 ;荧光显微镜及透射电镜下观察亲本细胞、敏感细胞及耐药细胞的超微结构。结果 :历时 6个月建立了卵巢癌TPT耐药细胞株A2 780 /TPT。此细胞株对TPT及米托蒽醌 (MX)耐药 ,耐药指数分别为 2 5及 19,对喜树碱 (CPT)轻微耐药 ,耐药指数为 3,对其他多种化疗药物不交叉耐药。耐药细胞的生长速度比亲本细胞慢 (P <0 .0 5 ) ,且细胞凋亡率及G2 M期细胞比例较亲本细胞高 (P <0 .0 5 )。敏感细胞呈典型的凋亡形态特征 ,而耐药细胞凋亡核的数量明显减少 ,凋亡程度明显减轻 ,且仍有较多的细胞核形态正常。结论 :所建立的卵巢癌TPT耐药株具有耐药细胞的基本生物学特征 ,且该耐药模型稳定。

Establishment of topotecan-resistant ovarian cancer cell line

Objective:To establish the topotecan (TPT)-resistant ovarian cancer cell line with its characteristics studied.Methods:The TPT-resistant cell line was established by repeatedly exposing ovarian epithelioma cell line A2780 to increased concentration of TPT.The resistance index of the resistant cell line to different chemotherapeutic drugs was determined by the methyl thiazolyl tetrazolium assay.Cell growth curve was painted by cell counting assay and the doubling time was accounted.Cell apoptosis rate and cycle distribution were measured by flow cytometry.The cell morphology of parental, sensitive and resistant cells were observed with fluorescence microscope and transmission microscope.Results:The TPT-resistant cell line A2780/TPT was established after 6 months and indicated drug-resistance to TPT and Mitoxantrone (MX),the drug resistance index being 25 and 19 respectively.The minor cross-resistance to camptothecin (CPT),the drug resistance index being 3, was tested in A2780/TPT cell line which still displayed sensitive to 5-FU,cDDP,MMC,taxol,ADM,VP-16 and MTX.The growing rate of resistant cells was obviously slower than that of the parental cells.The apoptosis rate and G 2M phase cell distribution of the resistant cells was significantly higher than that of the parental cells. The sensitive cells showed typical cell apoptosis characters but the amount and the degree of apoptosis nuclear in the resistant cells obviously decreased and the resistant cells still had many normal cell nuclears.Conclusion:TPT-resistant ovarian cancer cell line shows atypical resistant cell characters and the model of drug-resistance is steady.