卵巢癌紫杉醇耐药细胞耐药性和髓样分化因子88关系的研究  

Study on the relationship between drug resistance of Paclitaxel-resistant cell line in ovarian cancer and myeloid differentiation factor 88

作　　者：袁犁[1] 周琦[2] 徐发良[2] 李少林[1] 邹冬玲[2] 

机构地区：[1]重庆医科大学基础医学院放射医学教研室,重庆400016 [2]重庆市肿瘤研究所妇瘤科,重庆400030

出　　处：《重庆医科大学学报》2012年第7期570-574,共5页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(编号:81071812)

摘　　要：目的:研究卵巢癌亲本A2780细胞与卵巢癌紫杉醇耐药细胞A2780/Taxol生物学特性及与髓样分化因子(Myeloid dif-ferentiation factor 88,MyD88)的关系。方法:应用CCK-8法检测A2780细胞和A2780/Taxol细胞的耐药性和耐药指数,采用流式细胞法检测A2780和A2780/Taxol细胞的细胞周期。应用罗丹明123(Rhodamine 123,Rh123)排出实验测定P-糖蛋白(P-gly-coprotein,P-gp)表达。应用Western blot检测A2780和A2780/Taxol细胞中P-gp的表达情况。应用细胞免疫化学检测A2780和A2780/Taxol细胞中MyD88、P-gp的表达情况。结果:A2780与A2780/Taxol相比,A2780/Taxol生长缓慢,呈G1期阻滞;CCK-8结果显示A2780/Taxol细胞对紫杉醇的IC50为(36.81±2.05)μg/ml,A2780细胞对紫杉醇的IC50为(1.40±0.18)μg/ml;两者的耐药指数(Resistance index,RI)为26.35。Rh123排出实验显示A2780/Taxol平均荧光强度显著降低。Western blot检测发现A2780/Taxol的P-gp表达比A2780明显升高(P<0.05)。细胞免疫化学检测发现A2780/Taxol中的P-gp和MyD88表达明显高于A2780细胞。结论:A2780/Taxol细胞株具有明确的耐药性,紫杉醇耐药与MyD88的表达密切相关,该细胞株可用于卵巢癌紫杉醇耐药的基础研究。Objective：To investigate the biological characteristics of Paclitaxel-resistant cell line A2780/Taxol and parental cell A2780 of ovarian cancer and to discuss their relationship with myeloid differentiation factor 88（MyD88）.Methods：CCK-8 assay was applied to detect the drug resistance and resistance index of A2780/Taxol and A2780 cell.Flow cytometry was used to test the cell cycle of A2780 and A2780/Taxol.Rhodamine 123（Rh123） efflux was used and the expression of P-glycoprotein（P-gp） was evaluated.Western blot was used to detect the protein expressions of P-gp in A2780 and A2780/Taxol cell line.The expressions of MyD88 and P-gp were detected by immunocytochemistry.Results：Both cell lines had some changes in cell cycle.The G1 phase was longer in A2780/Taxol cell line compared with that in A2780 cell line,with statistical differences between the two groups（P〈0.05）.The results of CCK-8 showed that the IC50 of A2780/Taxol against Paclitaxel was（36.81±2.05） μg/ml while that of A2780 against Paclitaxel was（1.40±0.18） μg/ml.The resistance index of A2780/Taxol was 26.35.Mean fluorescence intensity in A2780/Taxol was significantly declined according to the results of Rh123 assay.The expressions of P-gp and MyD88 were significantly higher in A2780/Taxol cell line than in A2780 cell line detected by Western blot and immunocytochemistry.Conclusions：A2780/Taxol cell line is confirmed to be drug resistant.The expression of MyD88 is closely related with Paclitaxel-resistance,so it can be used for basic research of Paclitaxel-resistance.

关 键 词：紫杉醇耐药 卵巢癌 P-糖蛋白 髓样分化因子88 

分 类 号：R737.31[医药卫生—肿瘤]