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目的:建立人卵巢癌细胞株紫杉醇耐药细胞株OV1228/Taxol。方法:以人卵巢癌细胞株OV1228作为亲本细胞,采用浓度梯度递增法建立人卵巢癌紫杉醇耐药细胞株OV1228/Taxol。通过细胞形态学观察、生长曲线和群体倍增时间测定、药物敏感试验、细胞周期测定和罗丹明-123外排试验以及多药耐药基因(MDR1)和蛋白激酶C-α(PKC-α)基因及蛋白水平的测定,评价OV1228/Taxol的生物学特性。结果:成功建立起人卵巢癌紫杉醇耐药细胞株OV1228/Taxol,耐药指数为39.44,对顺铂(DDP)和5-氟尿嘧啶(5-FU)有交叉耐药性。与OV1228细胞相比,OV1228/Taxol耐药细胞异形明显,耐药细胞倍增时间显著延长,P=0.033 5;耐药细胞内罗丹明-123量显著减少,P=0.005 4;MDR1和PKC-α在基因和蛋白水平表达均明显增加。结论:OV1228/Taxol细胞对紫杉醇具有典型耐药特性,为进一步研究耐药逆转途径提供了实验基础。
Objective: To establish a ovarian cancer cell line paclitaxel resistant cell line OV1228 / Taxol. Methods: Human ovarian cancer cell line OV1228 was used as the parent cell, and the ovarian cancer cell line OV1228 / Taxol was established by increasing the concentration gradient. The results of cell morphology observation, growth curve and population doubling time, drug sensitivity test, cell cycle assay and rhodamine-123 efflux assay as well as multidrug resistance gene (MDR1) and protein kinase C-α And protein levels, evaluate the biological characteristics of OV1228 / Taxol. Results: The paclitaxel-resistant human ovarian cancer cell line OV1228 / Taxol was successfully established. The drug resistance index was 39.44. The resistance to cisplatin (DDP) and 5-fluorouracil (5-FU) was cross-resistant. Compared with OV1228 cells, the morphological changes of OV1228 / Taxol-resistant cells were obvious and the drug-resistant cell doubling time was significantly prolonged (P = 0.033 5). The amount of rhodamine-123 in drug-resistant cells was significantly decreased At the gene and protein levels were significantly increased. CONCLUSION: OV1228 / Taxol cells have typical drug resistance characteristics to paclitaxel and provide the experimental basis for further study of drug resistance reversal pathway.