大蒜素对体外人骨肉瘤细胞周期和细胞凋亡的影响

Effects of diallyl trisulfide on cell cycle and apoptosis of human osteosarcoma cell line Saos-2

目的:评价大蒜素(diallyl trisulfide)对人骨肉瘤细胞株Saos-2的增殖抑制效果,研究大蒜素对细胞周期和细胞凋亡的影响,并初步探讨其可能的分子作用机制。方法:采用25、50和100μmol/L大蒜素分别作用人骨肉瘤Saos-2细胞24、48、72和96h后,倒置显微镜下观察细胞形态学改变,MTT方法检测大蒜素对Saos-2细胞的增殖抑制效果,流式细胞仪检测细胞周期和细胞凋亡,并应用双向凝胶电泳和质谱分析技术分析Saos-2细胞中大蒜素敏感的蛋白质点。结果:大蒜素可在一定浓度范围内以浓度和时间依赖性的方式抑制Saos-2细胞增殖。大蒜素可使Saos-2细胞停滞在G0/G1期,并且诱导细胞凋亡,该作用也有时间和浓度依赖性。Saos-2细胞中有27个大蒜素敏感蛋白,其中下调18个,上调9个。结论:大蒜素可通过停滞细胞周期进程和(或)诱导细胞凋亡以达到抑制人骨肉瘤细胞株Saos-2细胞增殖的效果。蛋白质组学的研究结果初步提示了大蒜素抗肿瘤的分子作用机制,为今后研究骨肉瘤提供了新的作用靶点。

Objective： To investigate the effects of DATS （diallyl trisulfide） on cell proliferation, cell cycle and apoptosis of human osteosarcoma cell line Saos-2, and to explore their potential molecular mechanisms, l^ethods： Saos-2 cells were treated with 25, 50 and 100 μmol/L DATS for 24, 48, 72, and 96 h. The change of cell morphology was observed under an inverted microscope. The effect of DATS on the proliferation of Saos-2 cells was detected by MTT assay. After treatment with DATS at desired concentrations for various time intervals, the cell cycle and apoptotic rate of Saos-2 cells were analyzed by flow cytometry. The expressions of DATS-sensitive proteins in Saos-2 cells were systematically detected using 2-DE （two-dimensional gel electrophoresis） and mass spectrometry analysis. Results：DATS could inhibit the proliferation of Saos-2 cells in a dose- and time-dependent manner. Moreover, DATS could arrest the cell cycle at phase Go/G1 and induce the apoptosis in a time- and concentrationdependent manner. A total of 27 unique DATS-sensitive proteins, including 18 downregulated proteins and 9 upregulated proteins, were found in Saos-2 cells. Conclusion： DATS can suppress the proliferation of Saos-2 cells by blocking cell cycle progression and inducing apoptosis. The results of proteomics not only suggest the anticarcinogenic mechanism of DATS, but also provide a novel insight into the therapeutic targets for gene therapy of osteosarcoma.