摘要
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 taxol-resistance cell line of human ovarian carcinoma(OV1228/taxol),and investigate its biological features.METHODS:With the sensitive OV1228 human ovarian cancer cell line as the parental cells,taxol-resistant cell line of human ovarian cancer(OV1228/taxol) was established by continuous stepwise selection in the increasing concentration of taxol.Cell morphology,growth curve and population doubling time,drug sensitivity,cell cycle,cell uptake of Rh-123,as well as mRNA and protein levels of MDR1 and PKC-α were investigated to determine the biological features of OV1228/Taxol cell line.RESULTS: The resistance index(RI) of OV1228/Taxol cells was up to 39.44.They showed an obvious cross-resistance to DDP and 5-FU.Compared with OV1228 cells,OV1228/Taxol resistant cells were significantly heteromorphosis,and the population doubling time was significantly longer(P=0.033 5).The results of uptake of Rh-123 further proved that OV1228/Taxol cells had a significant resistance to Rh-123(P=0.005 4).PT-PCR and Western bolt studies showed that the levels of PKC-α and MDR1 mRNA and protein expression in the OV1228/Taxol cells were higher than those in the sensitive cells.CONCLUSION: OV1228/Taxol cell line shows typical multidrug resistance phenotypes and might serve as an ideal model for exploring the new route of reversing multidrug resistance.
出处
《中华肿瘤防治杂志》
CAS
2010年第23期1920-1923,共4页
Chinese Journal of Cancer Prevention and Treatment
基金
江苏省肿瘤医院重点课题(ZK200701)
江苏省卫生厅面上项目(H200842)
江苏省人事厅"六大人才高峰"项目资助(08-D-02)
关键词
卵巢肿瘤
紫杉酚
药物耐受性
细胞系
ovarian neoplasms
paclitaxel
drug tplerance
cell line