Spermidine/spermine N1-acetyltransferase gene expression in A549 cells treated with pomegranate juice

Introduction: Intracellular polyamines have an important role in cell proliferation and death pathway. Targeting polyamine metabolic pathway is therefore a potential intervention in cancers. Either downregulation of polyamines synthesis or induction of polyamine catabolism both had been shown to cause depletion of intracellular polyamines. Objective: This study was aimed to investigate the expression of genes involved in polyamine catabolism, spermidine/spermine N1-acetyltransferase (SSAT)in human lung carcinoma A549 cells after treated with pomegranate juice. Methods: A549 cells were grown in DMEM supplemented with 10% (v/v) fetal bovine serum and 1% (v/v) penicillin-streptomycin. For experiment, cells were seeded at 1×104 cell/ml and grown for 48 hour prior to treatment. A549 cells were treated with 2% pomegranate juice for various exposure times between 0 to 48 hours. RNA was isolated using Trizol reagent and the purity of RNA was measured using NanoDrop spectrophotometer. The assays were analyzed using two steps quantitative real-time PCR by using Bio-Rad CFX96 Real-Time PCR Systems. The assays were normalized against two reference genes which are GAPDH and β-actin. Results: In this study, pomegranate juice did not induce the expression of SSAT. After 0, 6, 12, and 24 hour exposure, the expression of SSAT remains unchanged (p>0.05) compared to control. At 48 hour exposure, the expression of SSAT also remains unchanged although the relative normalized expression was lowered compared to control (p