中国药理学通报
中國藥理學通報
중국약이학통보
CHINESE PHARMACOLOGICAL BULLETIN
2015年
4期
549-554
,共6页
乳腺癌%Salinomycin%阿霉素耐药%细胞凋亡%活性
乳腺癌%Salinomycin%阿黴素耐藥%細胞凋亡%活性
유선암%Salinomycin%아매소내약%세포조망%활성
breast cancer%salinomycin%doxorubicin resistance%cell apoptosis%ROS%mitochondrial mem-brane potential
目的：本研究旨在探讨 Salinomycin 对乳腺癌阿霉素耐药细胞株 MCF-7／DOX 增殖和凋亡的影响及可能作用机制。方法MTS 实验检测 Salinomycin 对 MCF-7／DOX 细胞增殖的影响；Annexin V-FITC ／PI 染色检测 Salinomycin 对MCF-7／DOX 耐药细胞凋亡的影响；DCFH-DA 染色检测 Sali-nomycin 对 MCF-7／DOX 耐药细胞活性氧（reactive oxygen species，ROS）产生的影响；JC-1法测定细胞线粒体膜电位；Western blot 法检测细胞凋亡相关蛋白 BAX、BCL-2、caspase-3和 caspase-9的表达变化。结果Salinomycin 能明显抑制MCF-7／DOX 耐药细胞增殖，且具有浓度依赖性；流式分析发现 Salinomycin 能够诱导 MCF-7／DOX 细胞凋亡，增加细胞内ROS 水平，降低细胞线粒体膜电位；与对照组相比较，Salino-mycin 处理明显抑制 BCL-2的表达，上调 BAX、cleaved caspase-3和 cleaved caspase-9的蛋白表达；抗氧化剂 N-ace-tylcysteine（NAC）则逆转上述作用。结论Salinomycin 能够诱导 MCF-7／DOX 细胞凋亡，其机制可能与 Salinomycin 诱导ROS 的产生，激活线粒体凋亡途径有关。
目的：本研究旨在探討 Salinomycin 對乳腺癌阿黴素耐藥細胞株 MCF-7／DOX 增殖和凋亡的影響及可能作用機製。方法MTS 實驗檢測 Salinomycin 對 MCF-7／DOX 細胞增殖的影響；Annexin V-FITC ／PI 染色檢測 Salinomycin 對MCF-7／DOX 耐藥細胞凋亡的影響；DCFH-DA 染色檢測 Sali-nomycin 對 MCF-7／DOX 耐藥細胞活性氧（reactive oxygen species，ROS）產生的影響；JC-1法測定細胞線粒體膜電位；Western blot 法檢測細胞凋亡相關蛋白 BAX、BCL-2、caspase-3和 caspase-9的錶達變化。結果Salinomycin 能明顯抑製MCF-7／DOX 耐藥細胞增殖，且具有濃度依賴性；流式分析髮現 Salinomycin 能夠誘導 MCF-7／DOX 細胞凋亡，增加細胞內ROS 水平，降低細胞線粒體膜電位；與對照組相比較，Salino-mycin 處理明顯抑製 BCL-2的錶達，上調 BAX、cleaved caspase-3和 cleaved caspase-9的蛋白錶達；抗氧化劑 N-ace-tylcysteine（NAC）則逆轉上述作用。結論Salinomycin 能夠誘導 MCF-7／DOX 細胞凋亡，其機製可能與 Salinomycin 誘導ROS 的產生，激活線粒體凋亡途徑有關。
목적：본연구지재탐토 Salinomycin 대유선암아매소내약세포주 MCF-7／DOX 증식화조망적영향급가능작용궤제。방법MTS 실험검측 Salinomycin 대 MCF-7／DOX 세포증식적영향；Annexin V-FITC ／PI 염색검측 Salinomycin 대MCF-7／DOX 내약세포조망적영향；DCFH-DA 염색검측 Sali-nomycin 대 MCF-7／DOX 내약세포활성양（reactive oxygen species，ROS）산생적영향；JC-1법측정세포선립체막전위；Western blot 법검측세포조망상관단백 BAX、BCL-2、caspase-3화 caspase-9적표체변화。결과Salinomycin 능명현억제MCF-7／DOX 내약세포증식，차구유농도의뢰성；류식분석발현 Salinomycin 능구유도 MCF-7／DOX 세포조망，증가세포내ROS 수평，강저세포선립체막전위；여대조조상비교，Salino-mycin 처리명현억제 BCL-2적표체，상조 BAX、cleaved caspase-3화 cleaved caspase-9적단백표체；항양화제 N-ace-tylcysteine（NAC）칙역전상술작용。결론Salinomycin 능구유도 MCF-7／DOX 세포조망，기궤제가능여 Salinomycin 유도ROS 적산생，격활선립체조망도경유관。
Aim To investigate the anti-proliferative effect of salinomycin on doxorubicin-resistant human breast cancer MCF-7 /DOX cells.Methods MCF-7 and MCF-7 /DOX cells were treated or untreated with salinomycin.Cell viability was detected by MTS assay. Cell apoptosis was detected by Annexin V-FITC /PI as-say.Reactive oxygen species (ROS)was measured by DCFH-DA staining.Mitochondrial membrane potential was measured by JC-1 assay.The expression of apopto-sis related proteins BAX, BCL-2, caspase-3, and caspase-9 were evaluated by Western blot analysis. Results The cell viability was significantly reduced by salinomycin treatment in a dose-dependent manner. The flow cytometry results showed that salinomycin in-duced MCF-7 /DOX cell apoptosis,increased ROS pro-duction,and decreased mitochondrial membrane poten-tial.Furthermore,salinomycin decreased the expres-sion of BCL-2,and increased the expression of BAX, cleaved caspase-3,and cleaved caspase-9.Moreover, the antioxidant N-acetylcysteine (NAC ) markedly blocked the above effects.Conclusions Our results suggest that salinomycin-induced apoptosis in MCF-7 /DOX is associated with induction of ROS production, and activation of mitochondria apoptosis pathway, which may become a potential chemotherapeutic agent for the therapy of doxorubicin resistant breast cancer.
