Anticancer effect of lactoferrin on Gastric Cancer Cell Line AGS

Abstract

Background: Lactoferrin is a glycoprotein with a molecular weight of 80 kDa, as a member of the transferring family. In recent investigations some important biological properties such as anti microbial, antiviral and anticancer activity of lactoferrin were studied. In the present study, the effect of antitumor induced by lactoferin was evaluated on stomach cancer cell AGS. Materials and Methods: Bovine milk was collected after parturition. After isolation of fat and casein, the milk’s other proteins were separated by ammonium sulfate precipitation. Then, lactoferrin was purified using CM-Sephadex-C50 cation exchange chromatography by FPLC system. The purified protein was identified using SDS-PAGE electrophoresis. The concentration of lactoferrin from milk and colostrum by Bradford assay obtained 0.6 and 2 mg/ml, respectively. Stomach cancer cell line, AGS and normal cell lines, HEK-293 were cultured in 37 °C and 5 percent CO2. After appropriate cell growth, different concentrations of purified lactoferrin were added to cells and incubated for 20, 36 and 48 hours. The cell viability and inhibition of cell growth were determined by MTT assay, also apoptosis assay was evaluated using propidium iodide staining through flow cytometry. Results: Lactoferrin (500µg/ml) inhibited the AGS growth by 55, 71 and 80% during 20, 36 and 48 h, respectively but did not have notable inhibitory effect on normal cell lines. The apoptosis level of AGS in flow cytometry was 55 and 70% in 20 and 48 h, respectively and was 5% in normal cells. Conclusion: The isolated lactoferrin from bovine milk showed inhibitory effect on stomach cancer cell line whereas; it did not have any significant effect on normal cells.