摘要
目的观察白芦藜醇(resveratrol,Res)对骨肉瘤细胞U-2OS增殖和凋亡的影响,并探讨其可能的分子机制。方法体外培养人骨肉瘤细胞U-2OS,用不同浓度的Res(5、10、20、40、80μmol/L)作用不同时间(24、48、72 h)后,MTT法检测Res对U-2OS细胞增殖活力的影响;流式细胞术检测Res对细胞周期和凋亡的影响;应用Caspase-3活性检测试剂盒检测Caspase-3酶活性;Western blot检测细胞内Bax、Bcl-2和Caspase 3蛋白的表达水平。结果 Res作用24 h时,与空白对照组及DMSO组比较,20和40μmol/L Res组的细胞增殖抑制率、G0/G1和G2/M期的细胞比例、细胞凋亡率、Caspase-3酶活性显著升高(P<0.05),S期细胞比例显著降低(P<0.05);20μmol/L Res组U-2OS细胞中Bax和Caspase-3蛋白表达水平显著升高(P<0.05),而Bcl-2蛋白表达水平明显降低(P<0.05)。结论 Res通过诱导提高Caspase-3酶活性,上调促凋亡基因Bax、Caspase-3及下调抗凋亡基因Bcl-2蛋白的表达水平,从而抑制U-2OS细胞的增殖,促进其凋亡,为Res治疗骨肉瘤提供了实验依据。
Objective To observe the effect of resveratrol（Res）on proliferation and apoptosis of human osteosarcoma cell line U-2OS, and investigate the potential molecular mechanism. Methods U-2OS cells were cultured in vitro and treated with Res at various concentrations（5, 10, 20 and 40 μmol / L）for 24, 48 and 72 h separately. The proliferation of U-2OS cells was measured by MTT assay, while the cell cycle and apoptosis rate by flow cytometry, the capase-3 activity by capase-3 activity kit, and the expression levels of Bax, Bcl-2 and caspase-3 proteins by Western blot. Results The inhibiting rate of proliferation, proportion at G0/ G1 and G2/ M stages, apoptosis rate and caspase-3 activity of U-2OS cells after treatment with 20 and 40 μmol / L Res for 24 h as well as expression levels of Bax and caspase-3 in the cells after treatment with 20 μmol / L Res increased significantly as compared with those in blank control and DMSO groups（P〈0. 05）, while the proportion of cells at S stage after treatment with 20 and 40 μmol / L Res and expression level of Bcl-2after treatment with 20 μmol / L Res decreased significantly（P〈0. 05）. Conclusion Res inhibited the proliferation and promoted the apoptosis of U-2OS cells by increasing the caspase-3 activity, up-regulating the expressions of Bax and caspase-3 and down-regulating the expression of Bcl-2 protein, which provided an experimental basis for treatement of osteosarcoma with Res.
出处
《中国生物制品学杂志》
CAS
CSCD
2016年第1期39-44,共6页
Chinese Journal of Biologicals
