DHA逆转卵巢癌细胞A2780/T紫杉醇耐药的机制研究-东南大学学报医学版

DHA逆转卵巢癌细胞A2780/T紫杉醇耐药的机制研究

单位：1. 南京医科大学附属江苏省肿瘤医院妇瘤科, 江苏南京 210009;
2. 解放军南京军区总医院普外科, 江苏南京 210002

关键词：二十二碳六烯酸卵巢癌紫杉醇耐药 P-糖蛋白核转录因子-κB信号通路 p38 促分裂素源活化蛋白激酶

分类号：R737.31;R979.1

出版年·卷·期（页码）：2015·34·第五期（689-695）

摘要：

目的:探讨二十二碳六烯酸(DHA)对卵巢癌耐药细胞的耐药逆转作用及其机制。方法:利用MTT评估细胞对紫杉醇耐药性的改变,罗单明实验验证细胞的药物外排能力,流式细胞仪检测细胞周期的分布,实时定量PCR及蛋白质印迹检测多药耐药蛋白1(MDR1)的mRNA水平、耐药相关蛋白及通路蛋白的蛋白水平。结果:DHA作用48 h后A2780/T对紫杉醇的半数抑制率(IC₅₀)下降(P<0.05),且罗丹明在胞内的含量增加,呈剂量依赖性(P<0.05)。同时紫杉醇与DHA联合使用可以改变细胞周期的分布,G_O/G₁期的细胞百分比上升(P<0.05),P-糖蛋白(P-gp)及其他MDR相关蛋白的表达下降,NF-κB及磷酸化p38 MAPK表达下降,而p38 MAPK未见明显变化。结论:DHA可以通过改变细胞周期分布,抑制P-gp及MDR相关蛋白的功能和表达,逆转卵巢癌耐药细胞的耐药性,同时该机制还可能与抑制NF-κB通路、抑制p38 MAPK的磷酸化有关。

Objective: To study the reversal effect of DHA on reversing drug resistance of taxol resistant ovarian cancer cells and to explore the underlying mechanisms in ovarian cancer cells. Methods: Using the MTT assay,the change of A2780/T cells resistant to taxol was determined. Drug efflux capacity in A2780/T cells was examined by flow cytometry using Rhodamine123(Rh123) intracellular accumulation assay. Cell cycle distribution was analyzed by flow cytometry(FCM). Real-time PCR was used to detect expression of MDR1 mRNA. Expression of drug resistance related proteins and proteins involved in the mechanisms was examined by Western bloting. Results: DHA significantly reduced the values of IC₅₀ of A2780/T cells to taxol(P<0.05). Additionally, Rh123 intracellular accumulation assay demonstrated that DHA promoted the chemotherapeutic drug accumulation to exert the cytotoxic effect on A2780/T cells in a concentration dependent manner (P<0.05).Our results also showed that taxol combined with DHA could change the cell cycle distribution and prolong the cell cycle in G₀/G₁ phase (P<0.05).Furthermore, we found that DHA down-regulated MDR related proteins, and suppressed the activation of NF-κB and phosphorylation of p38 MAPK. Conclusion: DHA reverses taxol resistance of A2780/T cells by changing the cell cycle distribution, inhibiting expression and function of P-gp and MDR related proteins, as well as blocking NF-κB and p38 MAPK pathways.

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