Molecular mechanism of Andrographolide on MMP-9 expression in human lung cancer cells / 中国生化药物杂志

ABSTRACT

Objective To investigate the effect and molecular mechanism of Andrographolide (AD) on matrix metalloproteinase-9 (MMP-9) expression in human colon cancer H 3255 cells. Methods Human lung cancer H3255 cell line weve cultured in vitro, and treated with 1.0, 3.0, 5.0μmol/L AD for 24 h, untreated cells was used as blank control. Cell viability, cell migration and cell invasion were analyzed by MTT assay, scratch healing assay and transwell membrane assay, respectably. Expression of MMP-9 mRNA was analyzed by RT-PCR. Protein expression and phosphorylation of Akt were detected by Western blot. Activity of NF-κB and MMP-9 were analyzed by luciferase reporter assay. Results AD could significantly reduced H 3255 cells invasion and migration without affecting the viability of cells, as demonstrated by scratch healing and transwell membrane assay. Furthermore, Western blot and RT-PCR results showed that AD could markedly inhibited MMP-9 activity and its expression in both protein and mRNA levels. AD could attenuated Akt’s phosphorylation and the activity of NF-κB. Moreover, LY 294002, an inhibitor of PI3 K, could significantly inhibited NF-κB transcriptional activity and MMP-9 expression. In addition, different concentrations of AD could inhibit the promote activity of MMP-9. Conclusion AD was a potential anti-invasive agent by inhibiting MMP-9 involved in PI3 K/NF-κB pathways.