Cathepsin L 在姜黄素诱导的胶质瘤细胞死亡中的作用
首发时间:2015-08-14
摘要:目的:以人脑胶质瘤U87和U251细胞为模型,探讨cathepsin L在姜黄素诱导的人脑胶质瘤细胞死亡中的作用及其机制。 方法:采用CCK-8法检测细胞活性;集落形成、Hoechst 33258荧光染色以及Annexin V/PI双染流式实验检测姜黄素对胶质瘤细胞U87和U251的细胞毒性作用。Western blot检测姜黄素对cathepsin L及cyclin B1和凋亡蛋白Bax/Bcl-2表达水平的影响,免疫荧光染色检测cathepsin L的变化。PI单染检测姜黄素对胶质瘤细胞周期的影响。用siRNA干扰cathepsin L观察姜黄素对细胞周期及细胞凋亡的影响。 结果:CCK-8、集落形成实验、Hoechst 33258荧光染色、流式细胞术及western blot结果显示,姜黄素能够诱导胶质瘤细胞的凋亡;PI单染流式细胞术结果表明姜黄素可以引起U87和U251细胞G2/M期阻滞。Western blot和免疫荧光法结果显示姜黄素能够激活cathepsin L。干扰 cathepsin L后,姜黄素诱导的细胞凋亡增加,细胞周期阻滞作用增强。 结论: Cathepsin L参与了姜黄素诱导的胶质瘤细胞凋亡和细胞周期阻滞。抑制cathepsin L可以增强胶质瘤细胞对姜黄素的敏感性。
关键词: Cathepsin L 姜黄素 胶质瘤细胞 凋亡 周期
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Cathepsin L involved in Curcumin-induced death of glioma cell
Abstract:Aim: To investigate the role of cathepsin L on curcumin-induced death in glioma cells.Methods:Cell Counting Kit (CCK)-8 assay, clonogenic survival assay, apoptosis assay ,Western blotting and cell cycle analysis were used to evaluate the effects of curcumin on cell proliferation, apoptosis, and the cell cycle. The expression of cathepsin L ,cyclin B1, bax/ bcl-2 was examined using Western blotting. The expression of cathepsin L also examined by immunocytochemistry. U87 and U251 cells were transfected with cathepsin L siRNA,then treated with curcumin. The apoptotic cells and cell cycle distribution was examined. Results: CCK8 assay and clonogenic survival assay showed that curcumin inhibited U87 and U251 cell proliferation. Flow cytometry analysis showed that curcumin induces apoptosis and cell cycle arrest. Expression of cathepsin L in U87 and U251 cells pretreated with curcumin increased. Cathepsin L siRNA indicated that cathepsin L influenced apoptosis and cell cycle arrest induced by curcumin. Conclusion: Our results suggest that cathepsin L was involved in the apoptosis and cell cycle arrest induced by curcumin. Inhibition of cathepsin L can enhance the sensitivity of glioma cells to curcumin.
Keywords: Cathepsin L curcumin glioma cell apoptosis;cell cycle arrest
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No.4652109108626914****
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