芒柄花黄素对人膀胱癌细胞T739增殖凋亡影响及机制探讨.docVIP

芒柄花黄素对人膀胱癌细胞T739增殖凋亡影响及机制探讨 　　[摘要] 目的 观察芒柄花黄素对人膀胱癌细胞T739增殖与凋亡的影响及其机制探讨。 方法 分别采用0、15、30、60 μmol/L的芒柄花黄素作用T739细胞后，CCK8法检测T739细胞的增殖情况，流式细胞术检测凋亡率的变化，Hoechst 33258荧光染色法观察凋亡情况，Western blot检测GSK-3β、Axin、β-catenin蛋白含量变化。 结果 15、30、60 μmol/L的芒柄花黄素以浓度和时间依赖方式明显抑制T739细胞的增殖（P 0.05），且作用T739细胞48 h后凋亡率随药物剂量增加显著上升（P 0.05），呈浓度-效应关系。Hoechst33258染色显示T739细胞随药物浓度的升高，凋亡明显增多。Western blot法显示GSK-3β及Axin蛋白表达随药物浓度升高而增加（P 0.05），β-catenin蛋白水平则随药物浓度升高而降低（P 0.05），且均呈浓度-效应关系。 结论 芒柄花黄素对T739细胞有显著的增殖抑制和诱导凋亡作用，其机制可能与芒柄花黄素上调GSK-3β、Axin蛋白表达和降低β-catenin蛋白水平有关。 　　[关键词] 芒柄花黄素；T739；凋亡；GSK-3β；Axin；β-catenin 　　[中图分类号] R285.5 [文献标识码] A [文章编号] 1673-7210（2017）09（a）-0004-04 　　[Abstract] Objective To study the effect of formononetin on the proliferation and apoptosis of human bladder cancer cells T739 cells， and discuss the mechanism. Methods Different contents of 0， 15， 30， 60 μmol/L formononetin were added to T739 cells. CCK8 assay was applied to determine the effect of formononetin on proliferation of T739 cells. Apoptosis was analyzed by flow cytometry and Hoechst33258 staining. The expressions of GSK-3β， Axin and β-catenin proteins in T739 cells were determined by Western Blot. Results Different contents of 15， 30， 60 μmol/L formononetin treatments suppressed the proliferation of T739 cells in a time- or dose-dependent manner （P 0.05）， and the apoptotic rates of formononetin-treated T739 cells for 48 hours were increased in a dose-dependent manner （P 0.05）. In addition， Hoechst 33258 staining revealed that the apoptotic cells of formononetin-treated T739 were increased in a dose-dependent manner. Further， Western blot results showed that formononetin significantly up-regulated the protein levels of GSK-3β and Axin in a dose-dependent manner （P 0.05）， while the protein level of β-catenin was inhibited （P 0.05）. Conclusion Formononetin exerts inhibitory growth effect and induced apoptosis on human bladder cancer of T739 cells. The underlyng mechanism involved may be associated with activating GSK-3β