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目的探讨姜黄素诱导人鼻咽癌NCE细胞凋亡的分子机制。方法通过吖啶橙-嗅化乙锭复合染色、原位末端脱氧核糖核苷酸转移酶分析法检测、电镜和DNA片段化分析法分析姜黄素对NCE细胞凋亡的影响,并应用流式细胞术、Western Blot、RT-PCR方法探讨姜黄素对细胞线粒体膜电位、半胱氨酸蛋白酶-3(cysteine proteases-3,caspase-3)酶活性、胞质CytC、Fas tuRNA及蛋白表达的影响。结果终浓度100μmol／L的姜黄素可诱导NCE细胞凋亡;与姜黄素共孵育12、24、48 h,凋亡细胞的阳性率分别为25．6％、40．3％、54．5％。姜黄素处理后NCE细胞线粒体膜电位下降,12、24、48 h的线粒体膜电位下降的阳性率分别是26．8％、42．3％、68．2％。caspase-3酶活性增强,12、24、48 h表达caspase-3酶活性细胞的阳性率分别是80．5％、100％、100％。胞质CytC含量显著增强,Fas mRNA及蛋白表达水平上升,Fas蛋白阳性率由33．6％上升到89．9％。结论姜黄素可能通过线粒体途径与死亡受体途径诱导鼻咽癌NCE细胞凋亡。
Objective To investigate the molecular mechanism of curcumin-induced apoptosis in human nasopharyngeal carcinoma cell line NCE. Methods Acridine orange - olfactory ethidium complex staining, in situ terminal deoxynucleotidyl transferase assay, electron microscopy and DNA fragmentation analysis of curcumin on NCE cell apoptosis, and the application of flow cytometry The effects of curcumin on mitochondrial membrane potential, cysteine protease-3 (caspase-3) activity, cytoplasmic CytC, Fas tuRNA and protein expression were investigated by Western Blot and RT-PCR. Results Curcumin at the final concentration of 100 μmol / L could induce apoptosis of NCE cells. The positive rates of apoptotic cells after curcumin incubation with curcumin for 12, 24 and 48 h were 25.6%, 40.3% and 54.5% . After treated with curcumin, the mitochondrial membrane potential of NCE cells decreased, and the positive rates of mitochondrial membrane potential decreased by 12.84 and 48 h were respectively 26.8%, 42.3% and 68.2%. The positive rate of caspase-3 activity was increased, and the positive rates of caspase-3 activity at 12,24 and 48 h were 80.5%, 100% and 100%, respectively. CytC content in cytoplasm increased significantly, Fas mRNA and protein expression increased, the positive rate of Fas protein increased from 33.6% to 89.9%. Conclusion Curcumin induces the apoptosis of nasopharyngeal carcinoma NCE cells through mitochondrial pathway and death receptor pathway.