应用细胞培养、细胞计数、流式细胞仪、琼脂糖凝胶电泳、Hoechst33258染色、苏木精-伊红(HE)染色、透射电镜观察等技术研究姜黄素对人成骨肉瘤MG.63细胞凋亡的诱导作用。实验结果显示,姜黄素可明显诱导MG-63细胞的凋亡,经7.5mg/L姜黄素诱导处理后,人成骨肉瘤MG-63细胞的增殖活动受到显著的抑制,细胞生长抑制率达63.14%;细胞周期检测出现亚二倍体(亚G1期)细胞峰值,细胞凋亡率达21.8%;琼脂糖凝胶电泳出现细胞凋亡典型的DNA“梯状”条带;细胞核经Hoechst33258染色出现浓染致密的固缩形态和颗粒状荧光:光镜和电镜观察结果显示,姜黄素处理组细胞体积缩小,细胞核固缩,染色质凝聚、线粒体肿胀、有凋亡小体形成等显著的凋亡特征。研究结果表明,姜黄素对人成骨肉瘤MG-63凋亡的诱导具有显著作用,从而为进一步研究细胞凋亡机制和多种骨骼疾病的研究提供重要基础和研究依据。
The apoptosis effects of curcumin (Cur) on human osteosarcoma MG-63 cells have been studied using the methods of cell culture, cell count, flow cytometry analysis, DNA gel electrophoresis, Hoechst33258 staining, HE staining, electron microscope. The results showed that proliferation of MG-63 cells could be inhibited by 7.5 mg/L Cur, and the inhibitory rate is 63.14%. The results of flow cytometry analysis showed that curcumin could induced the emergence of the phase of apoptosis, and the rate is 21.8%. Agarose gel electrophoresis revealed that cell DNA fragment exhibited characteristic "DNA ladder". Cell nucleus concentrated and appeared granular fluorescence by Hoechst33258 staining. Light microscope and electron microscope showed that the morphology of the cells treated with curcumin appeared shrinked, cell nucleus concentrated, chromatin agglutination, mitochondria swelling, and apoptosis body forming. This study suggested that curcumin had induced apoptosis of the human osteosarcoma MG-63 cells effectively, and provides important foundation and research proofs to study more about the apoptosis mechanisms of the osteosarcoma cells and the study of disease caused by osteoplast.