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吴茱萸碱(Evodiamine,Evo)对人结肠癌HCT-116细胞体外增殖和转移的影响及其可能机制初探。体外培养人结肠癌HCT-116细胞株,加入Evo1.5μmol/L、3μmol/L、6μmol/L、12μmol/L、24μmol/L、48μmol/L、96μmol/L继续培养48 h,CCK-8法检测Evo对HCT-116细胞生存活性的影响;Transwell法和划痕法检测STF-118804、Evo对HCT-116细胞体外迁移的影响;RT-PCR和Western blotting检测不同浓度的Evo(0μmol/L,1.5μmol/L,3μmol/L,6μmol/L)和STF-118804(10μg/L)诱导HCT-116细胞48 h后,细胞中SIRT1、NF-кB、MMP-9蛋白的表达情况。不同浓度的Evo(1.5μmol/L,3μmol/L,6μmol/L,12μmol/L,24μmol/L,48μmol/L,96μmol/L)作用于HCT-116细胞48 h后,细胞的增殖均受到明显抑制(p<0.05),且在一定范围内呈浓度依赖性;且Evo能抑制HCT-116细胞的迁移。Evo可明显下调HCT-116细胞内SIRT1、MMP-9 m RNA和蛋白的表达,上调NF-кB m RNA和蛋白的表达。体外实验表明,Evo对结肠癌细胞增殖和迁移的抑制作用是通过抑制SIRT1的表达,激活NF-кB信号通路,进而抑制MMP-9来实现的。
Effect of evodiamine on the proliferation and metastasis of human colon cancer HCT-116 cells in vitro and its possible mechanism. The human colon cancer HCT-116 cell line was cultured in vitro. The cells were cultured with Evo 1.5μmol / L, 3μmol / L, 6μmol / L, 12μmol / L, 24μmol / L, 48μmol / L and 96μmol / The effect of Evo on the survival of HCT-116 cells was detected by Transwell assay and scratch assay. The effects of STF-118804 and Evo on the migration of HCT-116 cells in vitro were detected by RT-PCR and Western blotting. The effects of Evo (0μmol / L, The expression of SIRT1, NF-κB and MMP-9 in HCT-116 cells induced by STF-118804 (10μg / L) for 48 h were detected by flow cytometry. Proliferation of HCT-116 cells was significantly inhibited by Evo (1.5μmol / L, 3μmol / L, 6μmol / L, 12μmol / L, 24μmol / L, 48μmol / L, 96μmol / L) Inhibition (p <0.05), and in a certain range of concentration-dependent; and Evo can inhibit HCT-116 cell migration. Evo significantly down-regulated the expression of SIRT1, MMP-9 mRNA and protein in HCT-116 cells and upregulated the expression of NF-κB m RNA and protein. In vitro experiments showed that Evo on colon cancer cell proliferation and migration inhibition by inhibiting the expression of SIRT1, activating NF-кB signaling pathway, thereby inhibiting MMP-9 to achieve.