木香烃内酯对人胆管癌RBE细胞生物学行为的影响及其作用机制

Effects of costunolide on the biological behaviours of cholangiocarcinoma RBE cells and its mechanism

目的:探讨木香烃内酯(costunolide,Cos)对人肝内胆管癌RBE细胞生物学行为的影响及其作用机制。方法:以不同质量浓度(0、2、4、6、8、12、16、20μg/ml)Cos作用RBE细胞,通过CCK-8法、流式细胞术分别检测Cos对细胞增殖能力和周期分布的影响,Annexin V-FITC/PI双染法、Transwell小室实验分别检测Cos对细胞凋亡和迁移侵袭的影响,q RT-PCR检测与侵袭相关因子MMP2、MMP9 m RNA的表达,Western blotting检测PI3K/AKT信号通路相关蛋白的表达。结果:Cos能够显著抑制RBE细胞的增殖活性(P<0.05或P<0.01),阻滞细胞周期于S、G2/M期,同时诱导RBE细胞的凋亡(P<0.01)、且呈质量浓度依赖性,Transwell及q RT-PCR结果显示Cos能够显著抑制RBE细胞的侵袭能力、降低侵袭相关转录因子MMP2和MMP9 m RNA的表达,Western blotting结果显示Cos明显抑制p-AKT、Bcl-2、MMP2及MMP9的表达,但促进Bax的表达。结论:Cos通过抑制PI3K/AKT信号通路降低胆管癌RBE细胞的增殖、迁移及侵袭能力。

Objective： To investigate the effects of costunolide （Cos） on the proliferation, apoptosis, migration andinvasion of cholangiocarcinoma RBE cells, and explore its potential mechanism. Methods： The CCK-8, flow cytometry, Annexin V-FITC/PI double staining,Transwell assays were used to examine the influence of Cos on proliferation, cell cycle, apoptosis, migration and invasion of RBE cells after treated with gradient concentrations of Cos. The expressions of MMP2 and MMP9 were detected by qRT-PCR, and the expression of PI3K/AKT-associated signal proteins was detected by Western blotting. Results： Cosdose-dependently inhibited proliferation activity of RBE cells（P〈0.05 or P〈0.01）, arrested cell cycle at S and G2/M phases and induced RBE cell apoptosis（P〈0.01）. Transwell and qRT-PCR results demonstrated that Cos impeded RBE cell migration, invasion, and reduced the transcription of MMP2 and MMP9. Cos inhibited the expressionofp-AKT, Bcl-2, MMP2 and MMP9, the level of Bax. Conclusion： Cos restrained the proliferation,migration and invasion of RBE cells by suppressing PI3K/AKTpathway.