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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。
Objective: To investigate the effect of tanshinoneⅡA (TanⅡA) on CNE and its possible mechanism. Methods: The effects of TanⅡA on the proliferation of CNE cells were observed by cell morphology, growth curve, MTT assay and cloning experiments. The effect of Tan Ⅱ A on CNE cell apoptosis was observed by Hoechest 33258 and PI double staining. The fluorescence intensity of TanⅡA On the calcium and mitochondrial membrane potential of CNE cells. The effect of TanⅡA on the expression of Bad and MT-1A mRNA in CNE cells was detected by RT-PCR. Results: Tan Ⅱ A inhibited the proliferation of CNE cells and increased with the increase of Tan Ⅱ A dose and the prolongation of action time. The IC50 of TanⅡA-treated CNE cells at 24, 48 and 72 h were 45.7, 24.8 and 3.3 mg.L -1, respectively. After treated with TanⅡA for 24 h, typical morphological changes of chromatin such as chromatin accumulation were observed in CNE cells. With the increase of TanⅡA dose, the percentage of CNE cells apoptosis gradually increased. After TanⅡA treatment, the intracellular calcium of CNE cells increased, the mitochondrial membrane potential decreased, the expression of Bad mRNA increased and the expression of MT-1A mRNA increased. CONCLUSION: TanⅡA has anti-CNE effect and its induction of apoptosis may be related to the upregulation of calcium-dependent pathway and MT-1A.