Fatemeh Amiri - Basic Sciences Group, Sari Agricultural Sciences and Natural Resources University, Sari, Mazandaran, Iran.
Fatemeh Moradian - Basic Sciences Group, Sari Agricultural Sciences and Natural Resources University, Sari, Mazandaran, Iran.
Alireza Rafiei - Molecular and Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Mazandaran, Iran

Background: Lactoferrin is a glycoprotein with a molecular weight of ۸۰ 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-C۵۰ 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 ۰.۶ and ۲ mg/ml, respectively. Stomach cancer cell line, AGS and normal cell lines, HEK-۲۹۳ and HFF were cultured in ۳۷ °C and ۵ percent CO۲. After appropriate cell growth, different concentrations of purified lactoferrin added to cells and incubated for ۲۰, ۳۶ and ۴۸ 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 ۵۰۰ µg/ml during ۲۰,۳۶ and ۴۸ h were ۵۵, ۷۱ and ۸۰percent , 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 ۵۵ and ۷۰ % in ۲۰ and ۴۸ 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.

Flowcytometry, Lctoferrin, MTT assay, Stomach cancer cell