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目的:探讨野菊花总黄酮对人骨肉瘤细胞增殖和凋亡的影响及其作用机制。方法:人骨肉瘤Saos-2细胞经野菊花总黄酮在不同浓度及不同时间作用后,倒置相差显微镜下观察细胞形态及细胞密度的变化,并采用CCK法检测野菊花总黄酮对细胞增殖的影响;流式细胞仪和逆转录-聚合酶链反应(RT-PCR)检测不同药物浓度作用细胞48 h后的凋亡率以及凋亡相关基因Caspase-3、BCL-2、BAX的表达情况。结果:野菊花总黄酮对Saos-2细胞的增殖有明显抑制作用,并呈剂量和时间依赖性,显微镜下观察可见细胞形态变圆,体积变小,内部颗粒增多等凋亡的形态变化。流式细胞仪检测细胞的凋亡率与药物浓度正相关。药物作用细胞48 h后,Caspase-3和BAX表达上调,BCL-2表达下降,BCL-2/BAX比率下降。结论:野菊花总黄酮对人骨肉瘤Saos-2细胞具有增殖抑制及诱导凋亡的作用,其机制可能与下调BCL-2/BAX比率而激活Caspase-3相关。
Objective: To investigate the effect of wild chrysanthemum total flavone on proliferation and apoptosis of human osteosarcoma cells and its mechanism. Methods: Human osteosarcoma Saos-2 cells were treated with different concentrations and time of total flavonoids of Chrysanthemum morifolium, the changes of cell morphology and cell density were observed under inverted phase contrast microscope. The effects of total flavonoids of Chrysanthemum morifolium on cell proliferation were detected by CCK method. Flow cytometry and reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the apoptosis rate and the expression of apoptosis-related genes Caspase-3, BCL-2 and BAX after 48 h treatment. Results: The total flavonoids of wild chrysanthemum significantly inhibited the proliferation of Saos-2 cells in a dose-and time-dependent manner. Morphological changes of apoptotic cells such as rounded cells, smaller volume and more internal granules were observed under the microscope. Flow cytometry detection of cell apoptosis rate and drug concentration is positively correlated. After 48 h, the expression of Caspase-3 and BAX were up-regulated, the expression of BCL-2 was decreased and the ratio of BCL-2 / BAX was decreased. CONCLUSION: The total flavonoids of Chrysanthemum indicum L. can inhibit proliferation and induce apoptosis of human osteosarcoma Saos-2 cells. The mechanism may be related to the down-regulation of BCL-2 / BAX ratio and activation of Caspase-3.