【摘要】 目的:探讨丹参酮ⅡA(tanshinoneⅡA,TanⅡA)抗人鼻咽癌CNE作用及其可能作用机制。方法:通过细胞形态学观察、生长曲线绘制、MTT检测以及克隆实验观察TanⅡA对CNE细胞增殖的影响;应用Hoechest33258和PI双染法观察TanⅡA对CNE细胞凋亡的影响;采用荧光分光光度计检测TanⅡA对CNE细胞细胞内钙及线粒体膜电位的影响;RT-PCR检测TanⅡA对CNE细胞Bad,MT-1A mRNA表达的影响。结果:TanⅡA能抑制CNE细胞增殖,且随TanⅡA剂量的增加和作用时间的延长而增强,TanⅡA作用CNE细胞24,48,72 h的IC50分别为45.7,24.8,3.3 mg.L-1。TanⅡA作用CNE细胞24 h后,CNE细胞出现染色质聚集等典型的凋亡形态学改变,且随TanⅡA剂量的增加,CNE细胞凋亡百分率逐渐增大。TanⅡA作用后,CNE细胞的细胞内钙升高、线粒体膜电位降低、Bad mRNA表达增加、MT-1A mRNA表达上调。结论:TanⅡA具有抗CNE作用,其诱导细胞凋亡可能与钙依赖性通路和MT-1A表达上调有关。更多还原

【Abstract】 Objective: To investigate anticancer effect and potential mechanism of tanshinone ⅡA(Tan ⅡA) on human nasopharyngeal carcinoma cell line CNE cells.Method: Antiproliferative effect of Tan ⅡA on CNE cells was evaluated by morphological examination,cell growth curves,colonial assay and MTT assay.Apoptosis detection was carried out using Hoechest33258 and PI double-dyeing method.Intracellular Ca2+ concentration and mitochondria membrane potential were detected by fluorospectrophotometer.Bad and MT-1A transcript analysis in CNE cells was analyzed by real-time reverse transcriptase-polymerase chain reaction(RT-PCR).Result: Tan ⅡA could inhibit CNE cells proliferation in dose-and time-dependent manner.50% inhibiting concentration of Tan ⅡA on CNE cells in 24,48,72 h was 45.7,24.8,3.3 mg·L-1,respectively.Typical apoptotic morphology such as chromatin aggregation was observed in CNE cells with Tan ⅡA treated for 24 h,and the apoptotic inducing effect was in a dose-dependent manner.After treated with Tan ⅡA,intracellular Ca2+ concentration of CNE cells was increased,mitochondria membrane potential of the cells was decreased,relative mRNA level of Bad and MT-1A was up-regulated.Conclusion: Tan ⅡA had anticancer effect on CNE cells through apoptosis via calcineurin-dependent pathway and MT-1A downregulation.更多还原