Objective To investigate the apoptosis-inducing effect of nitidine chloride in human osteosarcoma MG-63 cells and explore its mechanism. Methods The effect of nitidine chloride on the proliferation of MG-63 cells was detected by colorimetric MTT assay, and the morphological changes of cells treated with nitidine chloride were observed using fluorescence and electron microscope. Flow cytomerty was performed to analyze the apoptotic rate of the cells, and the protein expression levels of caspase-3, caspase-9, Bcl-2 and Bax were detected by Western blotting. Results Nitidine chloride inhibited the proliferation of MG-63 cells in a dose- and time-dependent manner. Fluorescence and electron microscopy revealed distinct apoptotic changes of the cells after nitidine chloride exposure. Flow cytometry indicated that nitidine chloride induced the apoptosis of MG-63 cells in a dose-dependent manner. Exposure to nitidine chloride, as shown by Western blotting, resulted in increased expressions of cleaved caspase-3, cleaved caspase-9 and Bax and decreased expressions of pro-caspase-3, pro-caspase-9 and Bcl-2. Conclusion Nitidine chloride can inhibit the proliferation of osteosarcoma cell line MG-63 by inducing cell apoptosis, the mechanism of which might be related with the activation of the caspase-dependent pathway.%目的 探讨氯化两面针碱对人骨肉瘤MG-63细胞诱导凋亡作用及其相关机制.方法 将不同浓度的氯化两面针碱作用于MG-63细胞,采用MTT法检测其增殖抑制作用;利用荧光显微镜和电镜观察其对细胞形态的影响;利用流式细胞仪检测不同浓度的氯化两面针碱诱导MG-63细胞的凋亡率;Westernblotting法分析caspase-3、caspase-9、Bcl-2和Bax表达的变化.结果 MTT结果表明,氯化两面针碱可明显抑制MG-63细胞的增殖,并存在时间-剂量依赖性.荧光显微镜下,氯化两面针碱处理过的细胞呈现典型的凋亡形态,如细胞核清亮,大小不一,有的裂解成亮蓝色的颗粒.透射电镜下,同样可看到细胞凋亡现象,如胞质内出现大量空泡,细胞核边集,细胞表面微绒毛减少.流式细胞仪结果表明氯化两面针碱诱导细胞凋亡呈明显的剂量依赖性,随着药物浓度升高,凋亡率升高.Western blotting证实,氯化两面针碱能上调MG-63细胞中cleaved-caspase-3、cleaved-caspase-9和Bax的表达,同时下调pro-caspase-3、pro-caspase-9和Bcl-2的表达.结论 氯化两面针碱可放导人骨肉瘤MG-63细胞凋亡,具体机制可能与激活caspase凋亡通路有关.
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