CAPE downregulate Rheb gene expression in human gastric cancer AGS cell line

Background: Ras homologue enriched in brain (Rheb) is an upstream regulator of mTOR signaling pathway, which regulates the process of cell-growth, cell cycle progression, proliferation and differentiation. The mammalian target ofrapamycin complex 1 (mTORC1) and Phospholipase D1 (PLD1) has become interesting targets for cancer therapy through theirs influences on oncogenic signals PI3K and AKT. Aim: Since Rheb is an upstream key activator of mTORC1 and PLD1, we investigated the effect of caffeic acid phenethyl ester (CAPE) on Rheb gene expression in human gastric cancer AGS cell line.Methods: AGS cell line was cultured in DMEM medium supplemented with 10% heat inactivated fetal bovine serumcontaining 1% penicillin-streptomycin at 37°C in a humidified incubator with 5% CO2. The cells were treated with CAPE (30 μM) for 48 h, total RNA was extracted, Reverse transcription of total RNA with oligo-dT was performed by first-strand synthesis kit (Fermentas) and quantitative Real time RT-PCR was carried out using SYBR Green PCR master mix. Analysis of relative Rheb gene expression data was performed using method. Results: Our results demonstrated that CAPE downregulates Rheb gene expression at mRNA level in human gastric cancer AGS cell line. Treatment with CAPE for 48 h decreased the expression level of Rheb mRNA with approximately 83% inhibition at 30 μM as analyzed by q-Real-time PCR. Conclusion: Our results strongly suggest that CAPE downregulate expression of Rheb gene that may be responsible of anticancer activity of CAPE by reduction of mTORC1 and PLD1 activity in AGS cancer cell line.

کلیدواژه ها:

AGS ، CAPE ، Rheb ، q-Real time RT-PCR