CAJ | 학술논문

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，GO／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。
목적：탐토이십이탄륙희산（ DHA）대란소암내약세포적내약역전작용급기궤제。방법：이용MTT평고세포대자삼순내약성적개변，라단명실험험증세포적약물외배능력，류식세포의검측세포주기적분포，실시정량PCR급단백질인적검측다약내약단백1（ MDR1）적mRNA수평、내약상관단백급통로단백적단백수평。결과：DHA작용48 h후A2780／T대자삼순적반수억제솔（ IC50）하강（ P＜0.05），차라단명재포내적함량증가，정제량의뢰성（ P＜0.05）。동시자삼순여DHA연합사용가이개변세포주기적분포，GO／G1기적세포백분비상승（P＜0.05），P-당단백（P-gp）급기타MDR상관단백적표체하강，NF-κB급린산화p38 MAPK표체하강，이p38 MAPK미견명현변화。결론：DHA가이통과개변세포주기분포，억제P-gp급MDR상관단백적공능화표체，역전란소암내약세포적내약성，동시해궤제환가능여억제NF-κB통로、억제p38 MAPK적린산화유관。
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.