Rice bran phytic acid induced apoptosis through regulation of Bcl-2/Bax and p53 genes in HepG2 human hepatocellular carcinoma cells.

Atheer Abbas Al-Fatlawi,Md Irshad,M Moshahid Alam Rizvi,Anees Abbas Al-Fatlawi,Md Zafaryab,Ayaz Ahmad

doi:10.7314/apjcp.2014.15.8.3731

Atheer Abbas Al-Fatlawi, Md Irshad + Show 4 more

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https://doi.org/10.7314/apjcp.2014.15.8.3731

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Abstract

Phytic acid (PA) has been reported to have positive nutritional benefits and prevent cancer formation. This study investigated the anticancer activity of rice bran PA against hepatocellular carcinoma (HepG2) cells. Cytotoxicty of PA (0.5 to 4mM) was examined by MTT and LDH assays after 24 and 48 h treatment. Apoptotic activity was evaluated by expression analysis of apoptosis-regulatory genes [i.e. p53, Bcl-2, Bax, Caspase-3 and -9] by reverse transcriptase-PCR and DNA fragmentation assay. The results showed antioxidant activity of PA in Fe3+ reducing power assay (p ≤ 0.03). PA inhibited the growth of HepG2 cells in a concentration dependent manner (p ≤ 0.04). After 48h treatment, cell viability was recorded 84.7, 74.4, 65.6, 49.6, 36.0 and 23.8% in MTT assay and 92.6, 77.0%, 66.8%, 51.2, 40.3 and 32.3% in LDH assay at concentrations of 1, 1.5, 2.0, 2.5, 3.0, and 3.5mM, respectively. Hence, treatment of PA for 24h, recorded viability of cells 93.5, 88.6, 55.5, 34.6 and 24.4% in MTT assay and 94.2, 86.1%, 59.7%, 42.3 and 31.6%, in LDH assay at concentrations of 1, 2.2, 3.0, 3.6 and 4.0mM, respectively. PA treated HepG2 cells showed up-regulation of p53, Bax, Caspase-3 and -9, and down- regulation of Bcl-2 gene (p ≤ 0.01). At the IC50 (2.49 mM) of PA, the p53, Bax, Caspase-3 and-9 genes were up- regulated by 6.03, 7.37, 19.7 and 14.5 fold respectively. Also, the fragmented genomic DNA in PA treated cells provided evidence of apoptosis. Our study confirmed the biological activity of PA and demonstrated growth inhibition and induction of apoptosis in HepG2 cells with modulation of the expression of apoptosis-regulatory genes.

Highlights

IntroductionPhytic acid (PA) (inositol hexaphosphate, IP6) is a natural compound, present mostly in legumes including corn, soy beans, nuts, wheat bran and rice bran (Canan et al, 2011)
Phytic acid (PA) is a natural compound, present mostly in legumes including corn, soy beans, nuts, wheat bran and rice bran (Canan et al, 2011)
Antioxidant activity Antioxidant activity of PA was examined by two different method based on DPPH radicals scavenging and ferrous ions reducing activity

Summary

Introduction

Phytic acid (PA) (inositol hexaphosphate, IP6) is a natural compound, present mostly in legumes including corn, soy beans, nuts, wheat bran and rice bran (Canan et al, 2011). Rice bran has been extensively examined for its biological activities and reported to have antioxidant , anti-inflamatory activity, lowering the risk of cancer formation, prevent coronary hearth diseases and decreased cholesterol level (Islam et al, 2011; Henderson et al, 2012). Rice bran have high content of PA ranging from 5.94 to 6.09g/100g (Liu et al, 2005). PA has been known for its beneficial effects on human health, in the prevention of renal calculi, diabetes, cancer and parkinson’s disease (Lee et al, 2005; Saw et al, 2007; Xu et al, 2008). PA has been reported for antioxidant, anticarcinogenic and hypolipidemic properties (Norazalina et al, 2010; Kang et al, 2012).

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