槲皮素抑制骨肉瘤细胞MG-63增殖作用研究

Inhibiting Proliferation Effects of Quercetin on the Osteosarcoma Cell Line MG-63

目的探讨槲皮素抑制骨肉瘤细胞MG-63增殖及促进其凋亡作用。方法以0、20、40和80μmol/L槲皮素分别处理MG-63细胞24h、48h及72h,以MTT法检测细胞增殖活力,以40μmol/L槲皮素处理MG-63细胞48h,用流式细胞术(FCM)检测细胞凋亡;逆转录-定量聚合酶链式反应(RT-qPCR)及Western Blot检测caspase 3mRNA及蛋白表达。结果不同剂量槲皮素对MG-63细胞增殖活力有明显抑制作用,且在48h,各剂量槲皮素处理组细胞抑制率均明显低于正常对照组,且呈现明显的剂量依赖性,差异有统计学意义(P<0.05);FCM检测结果显示,40μmol/L槲皮素处理MG-63细胞48h后,凋亡率明显高于正常对照组,差异有统计学意义(P<0.05);RT-qPCR结果显示,槲皮素处理后MG-63细胞caspase 3 mRNA相对表达量明显高于正常对照组,差异有统计学意义(P<0.05);Western blot结果显示,槲皮素处理后,MG-63细胞caspase 3前体及裂解产物相对表达量均明显高于正常对照组,差异有统计学意义(P<0.05),且caspase 3裂解比例(caspase 3cleaved-/pro-)亦明显增加,差异有统计学意义(P<0.05)。结论槲皮素可刺激MG-63细胞转录和表达caspase 3,加速其裂解,进而抑制MG-63细胞增殖活力。

Objective To detect the inhibiting effects of quercetin on the proliferation of osteosarcoma cell line MG-63.Methods The MG-63 were treated with 0、20、40 and 80μmol/L quercetin for 24 h、48 hand 72 hand then quantified the proliferation activity by MTT.Flow cytometry was applied to detect the apotosis of MG-63.Reverse transcript quantity polymerase chain reaction and Western Blot were applied to evaluate the caspase 3 mRNA and protein expression.Results All doses of quercetin had obvious the inhibitory effect on the proliferation of MG-63 cells.At 48 h,the cell inhibition rate of each dose of quercetin treatment group was significantly lower than that in the normal control group,and showed a significant dose dependence（P〈0.05）.The results of FCM results showed that after 48 hwhen the MG-63 cells were treated with 40 mol/L quercetin,the apoptosis rate was significantly higher than that of the normal control group（P〈0.05）.The RT-qPCR results showed that the relative expression of caspase 3 mRNA in quercetin treatment group was significantly higher than that of the normal control group（P〈0.05）.Western blot results showed that after quercetin treatment,the relative expression of caspase 3 precursor and lysate in MG-63 cells was significantly higher than that of the normal control group（P〈0.05）and the proportion of caspase 3 fragmentation（caspase 3 cleaved-/pro-）was also significantly increased（P〈0.05）.