Green tea polyphenols affect invasiveness of human gastric MKN-28 cells by inhibition of LPS or TNF-α induced Matrix Metalloproteinase-9/2

Biochim Open. 2016 Nov 5;3:56-63. doi: 10.1016/j.biopen.2016.10.002. eCollection 2016 Dec.

Abstract

Several studies demonstrated a correlation between green tea consumption and a reduced cancer risk. Among different components, green tea polyphenols have been identified as molecules responsible for the beneficial effects showed by the green tea against oxidative stress and cell invasiveness. In this study, we investigated the effects of green tea polyphenol extracts (GTPs) in human gastric MKN-28 cell line. To this aim, we have first evaluated the effect of GTPs on oxidative stress induced cell injury. The pre-treatment with 10-4 M catechin equivalents of GTPs exerts a protective effect on xanthine-xanthine oxidase induced cell cytotoxicity, thus confirming the anti-oxidant properties of GTPs. The effect of GTPs was also extended to the invasive ability of MKN-28 cells stimulated with TNF-α or LPS, as pro-inflammatory factors. Migration and matrigel invasion assays demonstrated that GTPs exposure (10-6 M) prevents the increase in cell invasiveness induced by TNF-α or LPS. Finally, we have analyzed the effect of GTPs on the levels of Matrix Metalloproteinases (MMP)-9/2, whose expression is up-regulated by TNF-α or LPS. Our results indicated that the pre-treatment with GTPs was able to reduce MMP-9/2 expression at both protein and enzyme activity levels in the conditioned media of TNF-α or LPS stimulated MKN-28 cells. In conclusion, our results demonstrated that green tea polyphenol extract reduces the invasiveness of gastric MKN-28 cancer cells through the reduction of TNF-α or LPS induced MMP-9/2 up-regulation. Therefore, these data support the hypothesis that GTPs could exert a protective role against the metastatic process in gastric cancer.

Keywords: Cell migration; Cell invasion; DMEM, Dulbecco's Modified Eagles's Medium; DMSO, Dimethyl sulfoxide; ECM, Extracellular matrix; FBS, fetal bovine serum; GTPs, Green tea polyphenols extract; Green tea polyphenols; LPS, Lipopolysaccharide; MKN-28 gastric cancer cells; MMP-, Matrix metalloproteinase; Matrix Metalloproteinase-2 (MMP-2); Matrix Metalloproteinase-9 (MMP-9); PBS, Phosphate-buffer saline; ROS, Reactive Oxygen Species; TNF-α, Tumor necrosis factor α.