目的观察穿心莲内酯(Andrographolide,AD)对肺癌细胞基质金属蛋白酶9(matrix metalloproteinase-9,MMP-9)表达的影响及其分子机制。方法体外培养肺癌细胞系H3255,分别用1.0,3.0和5.0μmol/L AD处理细胞24h,未处理组为空白对照。伤口愈合实验和侵袭实验分别检测AD对H3255细胞迁移与侵袭的影响。RT-PCR检测MMP-9 mRNA的表达;Western blot检测MMP-9蛋白表达和Akt磷酸化。荧光素酶报告基因分析NF-κB和MMP-9的活性。结果伤口愈合实验和Transwell小室实验结果显示,AD能在不影响细胞活性的条件下显著抑制H3255细胞侵袭和迁移。同时,Western blot和RT-PCR结果证实AD能抑制MMP-9蛋白及mRNA表达。此外,AD能抑制Akt磷酸化以及NF-κB的转录活性。PI3K抑制剂LY294002处理后,NF-κB-luc活性以及MMP-9蛋白表达显著降低。此外,不同浓度AD处理能显著抑制MMP-9的启动子活性。结论 AD是一种潜在的抗肺癌细胞转移药物,它可能通过影响PI3K/NF-κB通路抑制MMP-9表达而发挥作用。 Objective To observe the effect of Andrographolide (AD) on the expression of matrix metalloproteinase-9 (MMP-9) in lung cancer cells and its molecular mechanism. Methods Lung cancer cell line H3255 was cultured in vitro. The cells were treated with 1.0, 3.0 and 5.0 μmol / L AD for 24 h respectively. The untreated group was blank control. The effects of AD on the migration and invasion of H3255 cells were detected by wound healing test and invasion assay. The expression of MMP-9 mRNA was detected by RT-PCR. The protein expression of MMP-9 and Akt phosphorylation were detected by Western blot. Luciferase reporter assay was used to analyze the activity of NF-κB and MMP-9. Results Wound healing experiments and Transwell chamber experiments showed that AD could significantly inhibit the invasion and migration of H3255 cells without affecting cell viability. Meanwhile, Western blot and RT-PCR confirmed that AD inhibited MMP-9 protein and mRNA expression. In addition, AD can inhibit Akt phosphorylation and NF-κB transcriptional activity. PI3K inhibitor LY294002 treatment, NF-κB-luc activity and MMP-9 protein expression was significantly reduced. In addition, AD treatment at different concentrations significantly inhibited the promoter activity of MMP-9. Conclusion AD is a potential drug against lung cancer metastasis and may play a role in inhibiting the expression of MMP-9 by affecting the PI3K / NF-κB pathway.