Anticancer effect of lactoferrin on Gastric Cancer Cell Line AGS

Background: Lactoferrin is a glycoprotein with a molecular weight of 80 kDa, as a member of the transferring family. In recent investigations the 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 were collected after parturition. After isolation of fat and casein, the other milk’s proteins were separated by ammonium sulfate precipitation.Next step, lactoferrin was purified using CM-Sephadex-C50 cation exchange chromatography by FPLC system.The purified protein identified by 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 and HFF were cultured in 37 °C and 5 percent CO2. After appropriate cell growth, different concentrations of purified lactoferrin 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 statining through flow cytometry. Results: The results indicated that more inhibitory effect of lactoferrin with 500 µg/ml during 20,36 and 48 h were 55, 71 and 80percent , respectively for AGS but had not notable inhibitory effect on normal cell lines. The apoptosis level of AGS in analysis with flow cytometry was indicated 55 and 70 % in 20 and 48 h, respectively but not effective in normal cells. Conclusion:The isolated lactoferrin from bovine milk had inhibitory effect on stomach cancer cell line whereas, it didn’t has observable effect on normal cells.