毛蕊异黄酮抑制胶质瘤细胞增殖的机制研究  

The inhibitory effect of calycosin on the proliferation of glioma cells

作　　者：王维[1] 张路阳[1] 赵子龙[1] 张东勇[1] 包义君[1] WANG Wei;ZHANG Luyang;ZHAO Zilong;ZHANG Dongyong;BAO Yijun(Department of Neurosurgery,the First Affiliated Hospital of China Medical University,Shenyang 110001,China)

机构地区：[1]中国医科大学附属第一医院,神经外科,辽宁沈阳110001

出　　处：《沈阳药科大学学报》2020年第8期725-729,755,共6页Journal of Shenyang Pharmaceutical University

基　　金：沈阳市科技计划项目(17-230-9-13)。

摘　　要：目的探讨毛蕊异黄酮对胶质瘤细胞增殖的抑制作用。方法通过MTT实验检测毛蕊异黄酮对胶质瘤细胞(U87和U251)增殖的影响。用流式细胞仪检测不同处理组对细胞周期的作用。通过Western blot实验和Real time PCR实验检测毛蕊异黄酮对CDK4/6的作用。Kinase glo实验检测毛蕊异黄酮对EGFR激酶的作用。Western blot实验明确毛蕊异黄酮对EGFR激酶的抑制作用。结果对细胞处理24 h后,10μmol·L^-1和20μmol·L^-1毛蕊异黄酮处理组对U87细胞增殖抑制率分别是(17.05±0.78)%和(53.25±11.91)%;对U251细胞增殖的抑制率分别为(19.86±3.11)%和(29.07±2.99)%。流式实验指出毛蕊异黄酮对U87细胞的G1-S期阻滞作用明显。进一步western blot和real time PCR实验指出毛蕊异黄酮可以通过抑制CDK4和CDK6蛋白和RNA的表达来抑制U87细胞的增殖。接下来的Kinase glo实验得出毛蕊异黄酮可以显著抑制EGFR的激酶活性,western blot实验印证了这一结果。结论毛蕊异黄酮可以通过抑制EGFR激酶活性进一步抑制细胞周期相关蛋白CDK4/6来抑制胶质瘤细胞的增殖。Objective To investigate the inhibitory effect of calycosin on the proliferation of glioma cells.Methods The proliferation of glioma cells(U87 and U251)was detected by MTT assay after treated by different concentrations of calycosin.Cells cycle were tested by flow cytometry.The effect of calycosin on CDK4/6 was examined by Western blot test and Real time PCR assay.The effect of calycosin on EGFR kinase was examined by Kinase glo and Western blot.Results U87 cells were treated with 10μmol·L^-1 and 20μmol·L^-1of calycosin,the inhibition of cell proliferation were(17.05±0.78)%and(53.25±11.91)%respectively.U251 cells were treated with 10μmol·L^-1 and 20μmol·L^-1 of calycosin,the inhibition of cell proliferation was(19.86±3.11)%and(29.07±2.99)%,respectively.Flow cytometry showed that the inhibitory effect of calycosin on the G1-S phase of U87 cells was obvious.Further detection indicated that calycosin could inhibit the proliferation of U87 cells by inhibiting the expression of CDK4 and CDK6.Kinase glo experiments showed that genistein could significantly inhibit the activity of EGFR kinase,and western blot experiments confirmed this result.Conclusion Calycosin can inhibit the proliferation of glioma cells by inhibiting the activity of EGFR kinase and further inhibiting cell cycle related protein CDK4/6.

关 键 词：毛蕊异黄酮 胶质瘤 表皮生长因子受体 增殖 

分 类 号：R96[医药卫生—药理学]