Mechanism Of Ginsenoside Octanoate Rh2-O Inhibiting Liver Cancer Invasion And Metastasis By Regulating The C-jun/COX2/PGE2 Pathway

BackgroundLiver cancer is a highly malignant disease,which patients often dead for high metastasis and recurrence.Therefore,controlling the metastasis of liver cancer is one of the keys to preventing and treating liver cancer and improving the survival rate of liver cancer patients.A large number of studies have shown that ginsenosides have strong anticancer activity.The low bioavailability of ginsenosides is the main reason affecting their clinical application.In previous experiments,we used Rh2 and caprylyl chloride to synthesize Rh2 octanoate(Rh2-O).We found that Rh2-O has a higher rate of cellular uptake and stronger in vivo and in vitro anti-cancer effects than Rh2.But the effect of Rh2-O on cancer metastasis has not been studied.Therefore,in this paper,a series of experiments were designed to explore the effect and mechanism of ginsenoside octanoate Rh2-O against cancer metastasis.ObjectiveWe investigate the effect and possible mechanism of ginsenoside octanoate Rh2-O on liver cancer metastasis.MethodsIn this paper,the effects in vitro of ginsenoside Rh2 and its octanoate Rh2-O on Huh-7 cell metastasis were investigaged.We used CCK8 experiments to detect the toxicity of drugs to cells,used wound healing assay to detect the migration capacity of cells,used the Transwell experiment to detect the invasion ability of cells,used Western Blot(WB)experiments to detect protein expression levels in Huh-7 cells.In vivo,C57BL/6J mice were used as research objects,and an animal model of tumor-bearing mouse subcutaneous transplantation tumors was established using H22 mouse hepatoma cells to explore the effects of Rh2 and Rh2-O on the invasion of H22-bearing mouse transplantation tumors and the expression of EMT-related proteins.used vernier calipers to measure the tumor surface area of H22-bearing mice,and used precision scales to weigh,tumors and organs of mice used WB experiments to detect protein expression levels in tumor tissues.To investigage the role of c-jun/COX2/PGE2 pathway in the effect of Rh2 and Rh2-O in vitro,We used LPS to stimulate the expression of COX-2 in Huh-7 cells,used exogenous addition of PGE2 to increase the concentration of PGE2 acting on Huh-7 cells,and used transfection of c-jun c DNA to overexpress c-jun in Huh-7 cells.We used enzyme-linked immunosorbent assay(ELISA)to detect PGE2 secretion levels in cells,used immunoprecipitation(IP)experiments to detect the level of c-jun acetylation in cells,used CCK8 experiments to detect the toxicity of drugs to cells,used wound healing assay to detect the migration capacity of cells,used the Transwell experiment to detect the invasion ability of cells,used Western Blot(WB)experiments to detect protein expression levels in Huh-7 cells.ResultsIn vitro experiments showed that both Rh2 and Rh2-O could inhibit Huh-7 cell migration,invasion ability,invasion and expression of EMT-related proteins,could inhibit the expression of COX-2 protein and the secretion of PGE2 in Huh-7 cells,and could promote the acetylation of transcription factor c-jun,which in turn promotes its degradation.After overexpression of c-jun in Huh-7 cells,the expression of COX-2 in cells was significantly increased.After the COX-2 agonist LPS acts on Huh-7 cells,its PGE2 secretion increases significantly.After LPS agonized COX-2and exogenous addition of PGE2,the inhibitory effect of Rh2 and Rh2-O on Huh-7cell metastasis was partially reversed.In vitro experiments showed that Rh2 and Rh2-O had an effect on the weight and organ index of H22 subcutaneous transplanted tumor-bearing mice,had long-term inhibitory effect on the tumor surface area of H22-bearing mice,and inhibited tumor size,tumor cell invasion and expression of EMT-related protein and COX-2 protein in H22-bearing mice.ConclusionRh2 and Rh2-O inhibit liver cancer metastasis by promoting c-jun acetylation down-regulation of COX-2/PGE2 pathway.This article provides an experimental basis for the study of ginsenosides and their derivatives against tumor metastasis,provides scientific data for the functional exploration and application of ginsenosides.