Evaluation Of Proliferation Inhibition Effect Of Human Calprotectin On Human Gastric Cancer Cell Line (AGS) In Vitro

Document Type : Original Article

Authors

NATIONAL CELL BANK OF IRAN, PASTEUR INSTITUTE OF IRAN, TEHRAN, IRAN

Abstract

Introduction: Human calprotectin is an abundant protein in the neutrophil, monocyte and macrophage cytosol. Effect of proliferation inhibition of this protein has been elucidated on hepatoma, melanoma, murine fibrosarcoma lukemia and human adenocarcenoma breast cancer but not in human gastric cancer cell lines. In present study, in vitro proliferation inhibition effect of calprotectin in different concentrations and intervals on AGS (tumoral) and HGF (normal) cell lines was evaluated.
Material and Methods: Calprotectin was purified from human neutrophil by chromatography methods. The human gastric adenocarcinoma cell line (AGS) was used. These cells were maintained in RPMI 1640 medium supplemented with 10% FCS. AGS cells (10000 cell per well) were exposed to different concentration of calprotectin (25, 50, 100, 200, 400 µg/ml) at 24, 48, 72 h. For evaluation of calprotectin on normal cells, human gingival fibroblast (HGF) was incubated with 70µg/ml of calprotectin(almost 2 time of LC50 evaluated in AGS after 48 h). In this study MTT assay was used for evaluation of proliferation inhibition effect of calprotectin on AGS cells.
Results: Viability of the cells with increasing of calprotectin concentration was decreased. All of the cells in 400 µg/ml of calprotectin after 24 h were dead. Although, toxicity of the cells with increasing of incubation time was increased. The most cytotoxicity effect was evaluated on 72 h .The LC50 of calprotectin for AGS cell at 24, 48 and 72 h, relife 96.78, 38.66 and 9.86 µg/ml was determined, respectively. Comparison between proliferation inhibition effect of calprotectin on AGS cell and HGF was showed that difference is not significant but more effect on AGS cell lines.
Conclusion: These results showed that in compare to normal cell (HGF), calprotectin has more cytotoxicity effect on cancer cells (AGS). Calprotectin can be candidate as an anti-gastric cancer drug.

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