青藤碱对胆管癌细胞的抑制作用

Inhibitory effects of sinomenine on cholangiocarcinoma cells

目的 研究青藤碱对胆管癌细胞的抑制作用,并探究其可能的分子机制。方法 通过试剂盒检测青藤碱对胆管癌细胞caspase-3活性的影响,Western blot检测青藤碱对胆管癌细胞中增殖蛋白及PI3K/Akt通路的影响。应用PI3K/Akt抑制剂LY294002,探究PI3K/Akt通路在抗胆管癌中发挥的作用。通过裸鼠皮下成瘤实验,观察青藤碱对体内胆管癌恶性生物学效应的影响,并检测肿瘤组织中增殖、凋亡及PI3K/Akt相关蛋白表达的变化。结果 青藤碱可使胆管癌细胞中caspase-3活性增强。青藤碱可剂量依赖性地降低胆管癌细胞中增殖蛋白的表达,并抑制PI3K/Akt通路。PI3K/Akt特异性抑制剂LY294002可抑制胆管癌细胞的增殖蛋白表达,并可升高凋亡蛋白表达;与单用青藤碱比较,LY294002联合青藤碱可抑制胆管癌细胞增殖能力,并促进其凋亡;随着青藤碱剂量增加,其抑制荷瘤生长作用加强,并可降低胆管癌荷瘤中相关增殖蛋白与PI3K/Akt通路蛋白表达,升高凋亡蛋白表达。结论 青藤碱通过PI3K/Akt信号通路抑制胆管癌细胞增殖并促进凋亡,从而在体内外发挥良好的抗癌活性。

AIM To investigate the inhibitory effect of sinomenine on cholangiocarcinoma cells and its possible molecular mechanism.METHODS To the cholangiocarcinoma cells,sinomenine had its effects on caspase-3 activity determined by kit,and its influence on the proliferation protein and PI3 K/Akt pathway investigated by Western blot.The PI3 K/Akt inhibitor LY294002 was used to explore the role of PI3 K/Akt pathway in cholangiocarcinoma.The effect of sinomenine on malignant biological cholangiocarcinoma was observed in vivo through the subcutaneous tumorigenesis experiment on nude mice,in terms of variations of proliferation,apoptosis and PI3 K/Akt-related protein expressions of the tumor tissues.RESULTS Sinomenine not only enhanced the activity of caspase-3,but also reduced the expressions of proliferative proteins and inhibited the PI3 K/Akt pathway dose-dependently in cholangiocarcinoma cells.The PI3 K/Akt-specific inhibitor LY294002 inhibited the expressions of proliferative proteins and increased the expressions of apoptotic proteins.Compared with the single use of sinomenine,its combination use with LY294002 inhibited the proliferation of cholangiocarcinoma cells and promoted their apoptosis.To the tumor-bearing growth,sinomenine dose-dependently enhanced the inhibition on it,decreased its expressions of related proliferative proteins and PI3 K/Akt pathway proteins,and increased its expressions of apoptotic proteins.CONCLUSION Sinomenine exerts good anticancer activity in vitro and in vivo due to its contribution to proliferation inhibition and apoptosis activation of cholangiocarcinoma cells via PI3 K/Akt signaling pathway.