Impact and mechanism of Loganin on epithelial mesenchymal transformation of gastric cancer cells

Abstract

Objective To explore impact and molecular mechanism of Loganin on epithelial mesenchymal transformation of gastric cancer cells. Methods Human gastric cancer cell lines AGS, BGC823 and SGC7901 were cultured routinely. The effect of Loganin on cells activity was tested by utilizing methyl thiazol tetrazolium (MTT) assay. Also Wound assay and Transwell assay were employed to detect the effect of Loganin on migration and invasion in BGC823 cells. Furthermore, real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) and Western blotting were used to examine the variations of associated genes and phosphatidylinositol 3 kinase (PI3K)-protein kinase B (Akt) signal pathway in BGC823 cells prior and post Loganin interventions. Results Results of MTT assay showed that the activity of BGC823 was lowest among AGS (0.748±0.051), BGC823 (0.581±0.042) and SGC7901 (0.412±0.037) (F=44.101, P<0.01); the expression level of PCNA mRNA and protein declined gradually with the increase dose of Loganin (F=36.298, 47.942, P<0.01). Compared with the control group, the ability of migration and invasion in BGC 823 cells decreased, and this changes were particularly distinct in high dose Loganin group (200 mg/L), which were more significant than in low dose group (50 mg/L) (migration: F=87.750, P<0.01; invasion: F=85.749, P<0.01). Results of qPCR and Western blotting showed that expressions of migration, invasion associated genes in BGC823 cells changes significantly compared with the control group, especially in high dose Loganin group (P<0.01). Results of qPCR and Western blotting illustrated that EMT related gene in BGC823 cells changes significantly compared with the control group, especially in high dose Loganin group (P<0.01). The activity of PI3K-Akt signal pathway declined significantly post Loganin intervention, and expression of p-PI3K, p-Akt decreased significantly, especially in high dose Loganin group (F=38.590, 63.563; P<0.01). Conclusion Loganin may inhibit EMT in gastric cancer cells via regulating PI3K-Akt signal pathway. Key words: Gastric cancer; Loganin; Invasion; Epithelial mesenchymal transformation; Phosphatidylinositol 3 kinase-protein kinase B signal pathways