PURPOSE: The main aim of the study was to evaluate in vitro and in vivo the anticancer and apoptotic effects of neochlorogenic acid in human gastric carcinoma cell death and the underlying mechanism of apoptosis induction, reactive oxygen species (ROS) production and loss of mitochondrial membrane potential (MMP), m-TOR/PI3K/AKT signalling pathway, cell migration and cell invasion suppression. METHODS: Fluorescence microscopy using DAPI and annexin V/PI staining in combination with flow cytometry was used to study the apoptotic effects induced by neochlorogenic acid on gastric cancer cells. The effects on ROS and MMP were studied by flow cytometry. Western blot assay was used to evaluate the effects of neochlorogenic acid on m-TOR/PI3/Akt signaling pathway. To examine the anti-cancer activity of neochlorogenic acid in vivo, we used the nude mice xenograft model. RESULTS: The results indicated that neochlorogenic acid exhibited an IC50 of 20 µM in these cells. The study also showed that apoptosis was due to loss of MMP and increased intracellular ROS production. Neochlorogenic acid downregulated the expression of key proteins of m-TOR/PI3/Akt signaling pathway. After 6 weeks of neochlorogenic acid administration to mice, the average tumor volumes and growth for the untreated control group were significantly higher than the treated groups. CONCLUSION: Based on these results, we propose that neochlorogenic acid can be a prospective anti-cancer therapeutic lead for the management of human gastric carcinoma.
PURPOSE: The main aim of the study was to evaluate in vitro and in vivo the anticancer and apoptotic effects of neochlorogenic acid in human gastric carcinoma cell death and the underlying mechanism of apoptosis induction, reactive oxygen species (ROS) production and loss of mitochondrial membrane potential (MMP), m-TOR/PI3K/AKT signalling pathway, cell migration and cell invasion suppression. METHODS: Fluorescence microscopy using DAPI and annexin V/PI staining in combination with flow cytometry was used to study the apoptotic effects induced by neochlorogenic acid on gastric cancer cells. The effects on ROS and MMP were studied by flow cytometry. Western blot assay was used to evaluate the effects of neochlorogenic acid on m-TOR/PI3/Akt signaling pathway. To examine the anti-cancer activity of neochlorogenic acid in vivo, we used the nude mice xenograft model. RESULTS: The results indicated that neochlorogenic acid exhibited an IC50 of 20 µM in these cells. The study also showed that apoptosis was due to loss of MMP and increased intracellular ROS production. Neochlorogenic acid downregulated the expression of key proteins of m-TOR/PI3/Akt signaling pathway. After 6 weeks of neochlorogenic acid administration to mice, the average tumor volumes and growth for the untreated control group were significantly higher than the treated groups. CONCLUSION: Based on these results, we propose that neochlorogenic acid can be a prospective anti-cancer therapeutic lead for the management of human gastric carcinoma.