木犀草素上调miR-384对胶质瘤细胞增殖、侵袭和凋亡的影响Effects of luteolin on the up-regulation of miR-384 on proliferation, invasion and apoptosis of glioma cells
陈侠,来慧丽,赖满香,刘筱蔼,王海帆
CHEN Xia,LAI Huili,LAI Manxiang,LIU Xiaoai,WANG Haifan
摘要(Abstract):
目的 研究木犀草素(luteolin)对胶质瘤细胞增殖、侵袭和凋亡的作用及机制。方法 以质量浓度2.5、25和50μmol·L~(-1)Luteolin1处理U251和U87细胞。分别以克隆形成实验、流式细胞术、Transwell实验检测细胞克隆形成、凋亡和侵袭。采用Lipofectamine 2000将antagomir-NC、miR-384 antagomir转染胶质瘤细胞U251。以RT-qPCR检测胶质瘤细胞中miR-384的表达水平。转染miR-384 antagomir后用质量浓度50μmol·L~(-1)的luteolin处理,以蛋白免疫印迹法(western blot)检测各组细胞中Ki67、p21、VEGF、Bax、Bcl-2和N-cadherin蛋白相对表达水平。结果 结果表明CCK-8浓度>25μmol·L~(-1)时luteolin可显著降低U251和U87细胞活力;与对照组相比,以质量浓度25和50μmol·L~(-1) luteolin处理可明显降低细胞克隆形成率和侵袭率、提升细胞凋亡率。与miR-384 antagomir对胶质瘤细胞增殖、侵袭和凋亡的影响转染组比较,miR-384 antagomir+luteolin(50μmol·L~(-1))处理可显著下调Ki67、VEGF、Bcl-2和N-cadherin蛋白表达水平,上调p21和Bax蛋白表达水平。结论 木犀草素可通过上调miR-384水平抑制胶质瘤细胞增殖和侵袭能力并诱导其凋亡。
Objective Objective To study the effect of Luteolin on the proliferation, invasion and apoptosis of glioma cells and the relative mechanism.Methods Glioma cells U251 and U87 were treated with different concentrations of luteolin, and cell viability was detected by CCK8.U251 and U87 cells were treated with luteolin 12.5,25 and 50 μmol·L~(-1).Colony formation, apoptosis and invasion were detected by Colony formationtest, flow cytometry and transwell assay, respectively.Antagomir-NC and miR-384 antagomir were transfected into glioma cells U251 using lipofectamine 2000,and the expression level of miR-384 in glioma cells was assayed by RT-qPCR.After transfection with miR-384 antagomir, the cells were treated with luteolin 50 μmol·L~(-1),and the relative expression levels of Ki67,p21,VEGF,Bax, Bcl-2 and N-cadherin in each group were detected by Western blot.Results CCK-8 experiment results showed that Luteolin at the concentration greater than 25 μmol·L~(-1) can significantly reduce the viability of U251and U87 cells; compared with the control group, 25 μmol·L~(-1) and 50 μmol·L~(-1) luteolin significantly reduced the rate of cell colony formation and invasion, and increased the rate of cell apoptosis.Compared with miR-384 antagomir transfection group, miR-384 antagomir +luteolin(50 μmol·L~(-1)) treatment significantly down-regulated the protein expression levels of Ki67,VEGF,Bcl-2 and N-cadherin, up-regulated the protein expression levels of p21 and bax.Conclusion Luteolin can inhibit the proliferation and invasion of glioma cells and induce their apoptosis by up-regulating the level of miR-384.
关键词(KeyWords):
miR-384;胶质瘤;木犀草素;p21;VEGF;侵袭
miR-384;glioma;luteolin;p21;VEGF;invasion
基金项目(Foundation): 2020年度广东省医学科研基金资助项目(B2020212)
作者(Author):
陈侠,来慧丽,赖满香,刘筱蔼,王海帆
CHEN Xia,LAI Huili,LAI Manxiang,LIU Xiaoai,WANG Haifan
DOI: 10.14066/j.cnki.cn21-1349/r.2021.0301
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