苦杏仁苷抑制人肺癌NCI-H1299细胞体外侵袭的机制

Inhibitions and mechanisms of amygdalin in non-small cell lung cancer NCIH1299 cell line invasion in vitro

探讨天然产物苦杏仁苷对人非小细胞肺癌NCI-H1299细胞体外侵袭与转移的抑制作用及其机制。采用MTS法检测苦杏仁苷对肿瘤细胞生长的影响;采用Transwell小室和划痕试验检测苦杏仁苷对细胞侵袭和迁移能力的影响;采用Western blot和RT-PCR试验检测苦杏仁苷对MMP-2/9、integrinβ1、integrinβ4、整合素连接激酶(ILK)、黏附斑激酶(FAK)、p-FAK和β-catenin的表达水平,以及Akt和RICTOR的磷酸化水平的影响。结果表明:苦杏仁苷可显著抑制H1299细胞体外增殖,0.5和1 mg/m L苦杏仁苷作用H1299细胞48 h后,肿瘤细胞侵袭、迁移能力显著下降,MMP-2/9、integrinβ1/4、ILK、FAK、β-catenin蛋白和mRNA表达,MMP-2/9活性以及FAK、Akt和RICTOR磷酸化水平显著下调(P均<0.05)。

The aim of this study was to investigate the effects and mechanisms of a nature product, amygdalin, in non-small cell lung cancer NCI-H1299 cell line invasion and migration in vitro. NCI-H1299 （H1299） cells were treated with amygdalin. MTS assay was employed to determine cell proliferation, transwell chamber and wound- healing assay were employed to determine invasion and migration in vitro, western blotting and RT-PCR assay were employed to determine the expression of integrin β1, integrin β4, integrin-linked kinase （ILK）, focal adhe- sion kinase （ FAK）, p-FAK and β-catenin, and the phosphorylation of Akt and RICTOR. Results showed that in vitro proliferation of H1299 cells were inhibited significantly after treated with O. 5 and 1 mg/mL amygdalin for 48 h. The invasion and migration potential was decreased. The protein and mRNA expression of MMP-2/9, inte- grin β1/4, ILK, FAK, β-catenin, the activity of MMP-2/9, the phosphorylation of FAK, Akt and RICTOR were all deereased significantly（ all of P 〈 0. 05）.