汉黄芩素调节恶性胶质瘤U87细胞增殖和凋亡并影响miR-128的表达
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国家自然科学基金项目(81172389)


Wogonin regulated cell proliferation and apoptosis,and repressed miR-128 expression in human glioblastoma cell line U87
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    摘要:

    目的:观察汉黄芩素(wogonin)对人胶质瘤细胞株U87增殖和凋亡的影响,并初步研究miR-128在汉黄芩素影响U87细胞系中的作用机制。方法:汉黄芩素(浓度分别为25-50和100 μmol/L)作用于U87细胞24 h和48 h后,分别用荧光显微镜观察U87细胞形态,MTT法检测细胞增殖活性,流式细胞术检测细胞周期和细胞凋亡。并采用qRT-PCR检测U87细胞中miR-128的表达。结果:汉黄芩素处理24 h和48 h后,U87细胞数量明显减少,细胞核固缩或裂解;MTT检测结果显示U87细胞的增殖受到汉黄芩素的抑制并且与药物的作用时间和剂量有关;流式细胞术检测显示汉黄芩素处理后的细胞其大量停留在S期,汉黄芩素作用胶质瘤细胞24 h后活细胞比率下降,细胞死亡率增加。汉黄芩素处理后的U87细胞中miR-128的表达量较对照组明显增高。结论:汉黄芩素能抑制U87细胞的增殖,促进其凋亡,其机制与miR-128的上调可能有密切的相关性。

    Abstract:

    Objective:To investigate the effects of wogonin on human glioblastoma cell lines U87,and the role of miR-128 in the mechanism of wogonin effect on U87 cells. Methods:U87 cells were treated by wogonin within three different concentrations(25,50,100 μmol/L) for 24 h,48 h,respectively. The morphological change of U87 cells were observed by using fluorescence microscope. The MTT was used on cell proliferation test. Flow cytometry(FCM) was performed to analyze the cell cycle and cell apoptosis. The qRT-PCR of miR-128 was detected from U87 cells after treated with or without wogonin. Results:Obvious appearances of cell apoptosis were observed under fluorescence microscope,including chromatin condensation and nuclear fragmentation. The analysis of MTT assay showed that the cell proliferation of U87 was inhibited by wogonin and associated with dose- and time-dependent manner. The FCM showed that the U87 cells treated by wogonin were arrest in S phase and the apoptosis ratio of U87 treated by wogonin was significantly increased after 24 h. The expression of miR-128 increased after being treated with wogonin. Conclusion:Wogonin induces glioblastoma cell U87 apoptosis and may restrain cell proliferation through miR-128.

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陈望昊,王慧博,刘 宁.汉黄芩素调节恶性胶质瘤U87细胞增殖和凋亡并影响miR-128的表达[J].南京医科大学学报(自然科学版),2013,(4):456-461

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  • 收稿日期:2012-12-10
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  • 在线发布日期: 2013-04-29
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