黄连素抑制直肠癌SW480细胞增殖的实验研究
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四川省中医药管理局青年基金项目;四川省卫计委科研课题


Effect of berberine on proliferation and apoptosis of SW480 rectal cancer cells
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    摘要:

    【摘要】目的 观察黄连素抑制直肠癌SW480细胞增殖的效果并探讨其作用机制。方法 治疗组用不同浓度的黄连素(0μmol/L、25μmol/L、50μmol/L、100μmol/L、150μmol/L、200μmol/L)干预SW480细胞株。MTT比色法观察最佳作用浓度及细胞增殖效果。结果显示最佳作用浓度为150μmol/L。后续实验治疗组SW480细胞用150μmol/L黄连素干预48小时, 对照组用RPMI1640培养基。流式细胞仪检测两组SW480细胞的细胞周期, Annexin V检测细胞凋亡。免疫印迹法(Western blot)分析治疗组和对照组中Livin、YKL40、AKT和Cyclin D1蛋白的表达。酶联免疫吸附法检测两组促血管生成素2(Ang2)的表达水平。结果 黄连素干预SW480细胞后对细胞增殖有明显抑制作用, SW480细胞周期被阻滞在G1期并伴有细胞凋亡比例明显增加。Livin、YKL40、AKT和Cyclin D1蛋白的表达水平明显降低, 并且黄连素可明显抑制YKL40/AKT/Cyclin D1信号通路和SW480细胞中Ang2的表达水平。结论 黄连素能有效抑制SW480细胞的增殖, 其作用机制包括下调YKL40/AKT/Cyclin D1信号通路从而使细胞周期被阻滞在G1期, 通过降低Livin蛋白的表达诱导细胞凋亡, 并可下调Ang2的表达抑制血管生成。

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    【Abstract】Objective To examine the antitumor effect of BBR on rectal cancer cell lines SW480 in vitro and explored the mechanism.Methods SW480 cell lines were treated with varying BBR concentrations (025μmol/L, 25μmol/L, 50μmol/L, 100μmol/L, 150μmol/L and 200μmol/L). MTT assay was adopted to observe the optimal conceration and cell proliferation. The treatment group SW480 cell was treated with 150μmol/L BBR, and the control group was treated with RPMI1640 medium. Cell cycle of SW480 cell lines was measured with flow cytometry and apoptpsis of SW480 cell was examined by the Annexin V apoptosis assay. The expression of Livin, Ykl40, AKT and Cyclin D1 in SW480 cells were determined by Western blot analysis. Linked immunosorbent assay(ELISA) was used to measure Angiopoietin2 (Ang2) expression level in SW480 cells.Results Cell proliferation in SW480 cell lines was inhibited. The associated with the cell cycle was arrested in G1 phase and BBRtreated SW480 cells showed a significantly greater proportion of apoptosis. The expression levels of Livin, Ykl40, AKT and Cyclin D1 were notably decreased and BBR significantly reduced Ang2 expression level in SW480 cells.Conclusion This results showed that proliferation of rectal cancer SW480 cells was reduced upon BBR treatment. The mechanism involves arresting cell cycle in G1 phase by supressing YKL40/AKT/ Cyclin D1 signaling pathway, inducing apoptosis through decreased expression of Livin, and inhibits angiogenesis via downregulation of Ang2

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  • 在线发布日期: 2018-03-05
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