五味子乙素对人尤文肉瘤细胞抑制增殖及促进凋亡作用及其机制

Schisandrin B induces growth inhibition and apoptosis of human Ewing sarcoma cells in vitro

目的探讨五味子乙素(SchB)对人尤文肉瘤细胞株A673的抑制增殖和促进凋亡的作用机制及其相关信号通路表达。方法体外培养人尤文肉瘤细胞株A673,采用噻唑蓝(MTT)法研究不同浓度(0、10、20、50、100μmol/L)SchB对尤文肉瘤细胞的增殖抑制作用,并采用集落形成实验观察其(0、5、10、15μmol/L)浓度的SchB对尤文肉瘤细胞集落形成的抑制作用。Westernblot实验分析(10、20、50μmol/L)浓度梯度下SchB调控B细胞淋巴瘤/白血病-2(bcl-2)、bcl-2相关X蛋白(bax)、p53、细胞周期蛋白(Cyclin)D1蛋白的表达。结果SchB有明显抑制人尤文肉瘤细胞增殖作用,随浓度增加增殖抑制率为18.22%、22.50%、34.43%、53.56%;并呈现出浓度和时间依赖效果,同时还有明显抑制其集落形成能力,集落个数分别是110.5±12.4、82.2±14.1、54.3±9.2、38.4±16.5。对尤文肉瘤细胞中的凋亡蛋白研究显示SchB具有明显的抑制bcl-2和CyclinD1表达,同时促进bax和p53蛋白表达的作用。结论SchB对人尤文肉瘤细胞有明显抑制增殖和促进凋亡作用,其具体机制与抑制bcl-2和CyclinD1表达,促进bax和p53蛋白表达有关。

Objective To investigate the effect of Schisandrin B (Sch B) on growth inhibition and apoptosis of human Ewing sarcoma cells in vitro, and to explore its possible mechanism. Methods Ewing sarcoma cell line A673 was treated with Sch B in vitro. The inhibitory effect of Sch B on the proliferation of Ewing sarcoma cells was studied by methyl thiazol tetrazolium (MTT) assay (0, 10, 20, 50, 100 μmol/L). Colony forming experiment was used to explore the inhibitory effect of Sch B on the colony formation of Ewing sarcoma cells (0, 5, 10, 15 μmol/L). The expression of B cell lymphoma/leukemia-2 (bcl-2), bcl-2 associated X protein (bax), p53 and Cyclin D1 protein was detected by Western blotting (10, 20, 50 μmol/L). Results The results showed that Sch B effectively inhibited cell growth and induced apoptosis of human Ewing sarcoma cell line A673 (18.22%, 22.50%, 34.43%, 53.56%) in a time- and dose-dependent fashion. Sch B also significantly inhibited the colony forming ability of Ewing sarcoma cells (110.5±12.4, 82.2±14.1, 54.3±9.2, 38.4±16.5). Sch B could significantly down-regulate the expression of bcl-2 and Cyclin D1n, and significantly up-regulate the expression of bax and p53. Conclusion Our findings demonstrate that Sch B can significantly inhibit the proliferation of Ewing sarcoma cell line A673, which may be related with the down-regulation of bcl-2 and Cyclin D1, and up-regulation of bax and p53.