Kapral Małgorzata, Parfiniewicz Beata, Strzałka-Mrozik Barbara, Zachacz Agnieszka, Weglarz Ludmiła
Department of Biochemistry, Medical University of Silesia, Sosnowiec, Poland.
Acta Pol Pharm. 2008 Nov-Dec;65(6):697-702.
Over the last years phytic acid, a hexaphosphorylated inositol (IP6) has attracted particular attention due to its anti-cancer activity, however, the molecular mechanisms of its action have not been elucidated, as yet. The aim of this study was to evaluate the influence of phytic acid on the expression of genes encoding p65 and p50 subunits of NF-kappaB and of its inhibitor IkappaBalpha in human colorectal cancer cell line Caco-2. A kinetic study of p65 and p50 subunits and IkappaBalpha mRNAs expression was performed on Caco-2 cells after treatment with 1, 2.5 and 5 mM IP6 for 1, 6, 12 and 24 h. Quantification of the genes expression was carried out using real time QRT-PCR technique. Treatment of cells with 5 mM IP6 resulted in a strong increase in IkappaBalpha expression at 6 h, 12 h and 24 h. The level of p65 transcript after 1 h was lower in the cells exposed to 1, 2.5, and 5 mM IP6 than in the control cells. The increase in transcriptional activity of p65 gene in response to 5 mM IP6 after 6 h and 12 h was observed. Cells treated for 24 h with 2.5 mM and 5 mM IP6 showed a significant decrease in expression of p65 gene. There were no quantitative changes in the p50 gene expression in the cells treated with IP6 compared to the control cells. High positive correlation between the expression of IkappaBalpha and p65 was detected. The results of this study suggest that IP6 primarily influences p65 and IkappaBalpha genes expression in colon cancer cells. Changes in transcriptional activities of IkappaBalpha and p65 depend on IP6 concentration and time of its action.
在过去几年中,植酸(一种六磷酸化肌醇,即IP6)因其抗癌活性而备受关注,然而,其作用的分子机制尚未阐明。本研究的目的是评估植酸对人结肠癌细胞系Caco-2中编码NF-κB的p65和p50亚基及其抑制剂IκBα的基因表达的影响。在用1 mM、2.5 mM和5 mM IP6处理1小时、6小时、12小时和24小时后,对Caco-2细胞进行p65和p50亚基以及IκBα mRNA表达的动力学研究。使用实时定量逆转录聚合酶链反应(QRT-PCR)技术对基因表达进行定量。用5 mM IP6处理细胞后,在6小时、12小时和24小时时IκBα表达显著增加。在暴露于1 mM、2.5 mM和5 mM IP6的细胞中,1小时后p65转录本水平低于对照细胞。在6小时和12小时后,观察到p65基因转录活性因5 mM IP6而增加。用2.5 mM和5 mM IP6处理24小时的细胞显示p65基因表达显著降低。与对照细胞相比,用IP6处理的细胞中p50基因表达没有定量变化。检测到IκBα和p65表达之间存在高度正相关。本研究结果表明,IP6主要影响结肠癌细胞中p65和IκBα基因的表达。IκBα和p65转录活性的变化取决于IP6浓度及其作用时间。