Effect of Saffron Extract on Expression of Bax and Bcl-2 Genes in Gastric Adenocarcinoma Cell Line (AGS)

Background: Stomach cancer is one of the most common cancers in modern societies. B-Cell Lymphoma-2 (Bcl-2) family members are classified to anti-apoptotic and pro-apoptotic groups, based on structural and functional features. The Bcl-2 inhibits apoptosis, while Bax (BCL-2 associated X protein) induces the apoptosis process. Saffron has anti-cancer properties and inhibits tumor genesis. The aim of this study was to investigate the anti-cancer properties of saffron extract and its effect on expression of Bcl-2 and Bax genes in gastric adenocarcinoma cell line (AGS) by real-time polymerase chain reaction (PCR). Methods: In this experiment, aqueous extract of saffron was made and MTT test was performed on AGS cells at concentrations of 0, 200, 400, 800, 1200, and2000g/mLfromsaffron extract at 48and72 hours. The50% inhibitory concentration (IC50)wascalculated for saffron extract at specific dosesandtimes. TheAGScancer cell linewastreated by saffron extract with concentrations of 800, 1200, and 2000 g/mL at 48 and 72 hours. Total RNA was extracted from AGS cells and its concentration and purity was determined. Then, cDNA was synthesized and evaluating the expression of Bcl-2 and Bax genes was performed by real-time PCR. Finally, the obtained results were analyzed by a statistical software. Results: The results of MTT test showed that with increasing concentration of extract and time, disappearance of cancer cells is increased. Thus, saffron extract has high cytotoxicity at high concentrations and in the long-term. The expression of Bax and Bcl-2 genes was also associated with significant increase (P < 0.05) in 800, 1200, and 2000 g/mL extract doses and at 48 and 72 hours, compared to cells that were not treated with the saffron extract. The overall result of this experiment showed that the ratio of Bax on Bcl-2 expression (Bax/Bcl-2) increased with increasing extract concentration and time. This results in the apoptosis of cancerous cells. Conclusions: Treatment of AGS cells with saffron extract caused an increase in the expression rate of Bax gene in comparison with Bcl-2 gene, and AGS cells were driven towards cell death. According to the obtained results, this effect is dependent on dose and time.