染料木黄酮通过PI3K/AKT途径抑制雄激素非依赖性LNCaP细胞 |
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引用本文: | 曹玉霖,李飞,赵欢,欧愚,张薇薇,朱彦锋. 染料木黄酮通过PI3K/AKT途径抑制雄激素非依赖性LNCaP细胞[J]. 安徽医科大学学报, 2018, 53(7): 1017-1021. DOI: 10.19405/j.cnki.issn1000-1492.2018.07.006 |
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作者姓名: | 曹玉霖 李飞 赵欢 欧愚 张薇薇 朱彦锋 |
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作者单位: | 成都医学院公共卫生系,成都,610500;南充市卫生学校教务科,南充,637000 |
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基金项目: | 国家自然科学基金(81402678),四川省卫生和计划生育委员会科研课题(17PJ442),成都医学院国家级大学生创新实验项目(201613705003、201713705031) |
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摘 要: | 目的 利用LNCaP细胞建立去势抵抗前列腺癌细胞模型,研究染料木黄酮(GEN)对去势抵抗前列腺癌细胞生长的影响及可能的机制.方法 在无激素的血清环境下长时间培养LNCaP细胞,获得稳定生长的雄激素非依赖性 LN-CaP前列腺癌细胞,并鉴定细胞.以不同浓度的 GEN(0、6. 25、12. 5、25、50、100、200 μmol/L)分别作用于雄激素依赖性LNCaP和雄激素非依赖性LNCaP细胞24、48、72 h ,利用CCK-8法检测细胞增殖活性,Western blot法检测前列腺特异性抗原(PSA)、P53抑癌基因、细胞周期蛋白(Cyclin D1)、增殖细胞核抗原( PCNA)在雄激素非依赖LNCaP细胞中的表达水平. Western blot法检测其对PI3K/AKT信号通路的调节作用.结果 GEN对雄激素非依赖性LNCaP细胞增殖具有明显的抑制作用,并呈剂量、时间依赖关系. Western blot显示雄激素非依赖性前列腺癌细胞内PCNA、Cyclin D1随着GEN浓度的增加逐渐下调,P53随着GEN浓度的增加逐渐上调(P<0. 05). GEN抑制了PI3K和AKT的磷酸化.结论 GEN可以在体外抑制雄激素非依赖性LNCaP细胞的增殖,其作用可能是通过抑制PI3K/AKT信号通路,进而抑制细胞增殖实现的.
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关 键 词: | 染料木黄酮 去势抵抗前列腺癌 增殖 雄激素非依赖性 信号通路 |
Geinsten inhibits androgen independent LNCaP cells through the PI3K/AKT pathway |
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Abstract: | Objective To explore the effect of genistein ( GEN) on castration-resistant prostate cancer ( CRPC) cells with an androgen independent LNCaP cells model. Methods An androgen independent LNCaP cell model was induced by culturing human prostate cancer LNCaP cell in hormone free medium for 40 generations, and then iden-tified by PSA expression. Androgen dependent and independent LNCaP cells were respectively treated with GEN at the concentrations of 0,6. 25, 12. 5, 25, 50, 100, 200 μmol/L for 24, 48 and 72 hours followed by proliferation test with CCK-8 assay. Expression of P53, Cyclin D1 and PCNA in the cells were detected by Western blot. The effect of GEN on PI3K pathway was determined by Western blot. Results GEN inhibited androgen independent LNCaP cell proliferation in dose and time-dependent manner. The expression of PCNA and Cyclin D1 were down-regulated, and that of P53 up-regulated with the increased concentration of GEN( P <0. 05) . GEN inhibited the phosphorylation of PI3K and AKT. Conclusion GEN inhibits the proliferation of androgen independent LNCaP cells by inhibiting PI3K/AKT signaling pathway. |
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