杨梅黄酮对小鼠脑胶质瘤GL261细胞增殖和凋亡的影响

目的：观察杨梅黄酮对小鼠脑胶质瘤GL261细胞增殖、凋亡、迁移和侵袭的影响，为杨梅黄酮应用于胶质瘤的治疗提供理论依据。方法：体外培养小鼠脑胶质瘤GL261细胞，实验分为对照组和不同浓度杨梅黄酮组，应用CCK-8法检测细胞的存活率，流式细胞术检测各组GL261细胞的细胞周期，Hoechst 33342染色及流式细胞术检测各组GL261细胞的细胞凋亡情况，应用Transwell小室检测各组GL261细胞迁移数目和进入小室下部的细胞数量，应用RT-PCR法检测杨梅黄酮处理前后GL261细胞中基质金属蛋白酶（MMPs）mRNA的表达水平。结果：CCK8法检测，与对照组比较，20 μmol·L^-1杨梅黄酮组细胞存活率明显下降（P<0.01）。细胞周期检测，与对照组比较，40 μmol·L^-1杨梅黄酮处理组GL261细胞G₁期比例升高（P<0.05），S期比例降低（P<0.05）。Hoechst 33342染色，杨梅黄酮组GL261细胞出现凋亡小体。流式细胞术AnnexinⅤ/PI荧光双染检测，与对照组比较，杨梅黄酮组细胞早期凋亡率和晚期凋亡率均增加（P<0.05）。Transwell小室迁移和侵袭实验，与对照组比较，5、10和20 μmol·L^-1杨梅黄酮组发生迁移的细胞数目明显减少（P<0.05）。Transwell小室检测，与对照组比较，杨梅黄酮组进入下室的细胞数目明显减少（P<0.05），且随着杨梅黄酮浓度的增加，穿过Matrigel的细胞数目逐渐减少。与对照组比较，5和10 μmol·L^-1杨梅黄酮组GL261细胞中MMP-2和MMP-9 mRNA表达水平均降低（P<0.05）。结论：杨梅黄酮可抑制胶质瘤细胞的增殖，诱导其凋亡，并抑制其迁移及侵袭。

Effects of myricetin on proliferation and apoptosis of glioma GL261 cells in mice

Objective:To observe the effects of myricetin on the proliferation, apoptosis, migration and invasion of mouse glioma GL261 cells, and to provide theoretical basis for myricetin in treating glioma. Methods:The mouse glioma GL261 cells were cultured in vitro. The experiment was divided into control group and different concentrations (10, 20, 30, 40, 50 and 60 μmol·L^-1) of mytricetin groups. CCK-8 assay was used to detect the survival rates of the mouse glioma GL261 cells in various groups.The cell cycle of GL261 cells in various groups was detected by flow cytometry; Hoechst 33342 staining and flow cytometry were used to detect the apoptosis of Gl261 cells in various groups. The number of migrated GL261 cells and the number of GL261 cells entering the lower chamber were detected by Transwell chamber assay. The expression levels of matrix metalloproteinases(MMPs) mRNA were detected by RT-PCR method. Results:The CCK-8 assay results showed that the survival rate of GL261 cells in 20 μmol·L^-1 myricetin group was significantly decreased compared with control group (P<0.01). The flow cytometry results showed that the proportion of GL261 cells in G₁ phase in 40 μmol·L^-1 myricetin group was significantly increased (P<0.05) and the proportion of GL261 cells in S phase was decreased(P<0.05) compared with control group.The Hoechst 33342 staining results showed that the apoptotic bodies appeared in myricetin groups.The early and late apoptotic rates of GL261 cells in myricetin groups detected by AnnexinⅤ/PI double staining were increased compared with control group(P<0.05). Compared with control group,the number of migrated GL261 cells in 5,10,and 20 μmol·L^-1 myricetin groups was decreased in Traswell chamber assay (P<0.05).Compared with control group,the number of GL261 cells entering the lower chamber in myricetin groups was decreased significantly(P<0.05);the number of GL261 cells passing the Matrigel was gradully decreased with the increasing of the concentrations of myricetin.Compared with control group,the expression levels of MMP-2 and MMP-9 mRNA in the GL261 cells in 5 and 10 μmol·L^-1 myricetin groups were decreased(P<0.05). Conclusion:Myricetin could inhibit the proliferation of GL261 cells and induce the apoptosis. Meanwhile, it could also inhibit the migration and invasion of GL261 cells.