Peptides extracted from common bean (Phaseolus vulgaris L.) non-digestible fraction caused differential gene expression of HCT116 and RKO human colorectal cancer cells
Peptides extracted from common bean (Phaseolus vulgaris L.) non-digestible fraction caused differential gene expression of HCT116 and RKO human colorectal cancer cells
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DOI：
10.1016/j.foodres.2014.02.037
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发表时间：
2014-08-01
影响因子：
8.1
通讯作者：
Gonzalez de Mejia, Elvira
中科院分区：
文献类型：
--
作者：
Luna Vital, Diego A.;Loarca-Pina, Guadalupe;Gonzalez de Mejia, Elvira
研究了普通菜豆（Phaseolus vulgaris L.）不明目的是通过微阵列分析，使用人结肠癌细胞来检查从普通菜豆Azufrado Higuera（AH-PE）和Bayo Madero（BM-PE）的两个栽培品种中提取的肽对基因表达的影响。用0.5mg/mL AH-PE和BM-PE处理HCT 116和RKO结肠癌细胞24小时;提取RNA，并使用Agilent低输入快速荧光标记测定法分析基因表达。在HCT 116人类细胞中，AH-PE和BM-PE处理分别影响了511和964个基因，其中发现405个基因通常受到肽提取物的影响。在RKO人细胞中，AH-PE和BM-PE处理分别显著影响了45和32个基因，两种提取物共有19个基因。利用数据库进行生物信息学分析表明，这些基因主要与氧化还原、刺激反应、蛋白磷酸酶、MAPK信号、硒结合和创伤反应等有关。抗氧化途径分析显示，在HCT 116细胞系中，NRF-2相关的抗氧化酶上调，而谷胱甘肽氧化还原相关的酶在RKO细胞系中下调，可能导致活性氧（ROS）失衡。通过RT-PCR对基因的验证显示出与在癌症增殖和凋亡中具有潜在作用的基因（PDE 4 B和OSGIN 1，以及KRT 19和EEF 1A 2）的良好一致性。该研究首次发现，两种豆类品种触发的两种人结肠癌细胞系中有8个差异表达基因相似，即C11 orf 31、C9 orf 169、EMP 1、GEM、PLIN 2、SUN 3、TRIM 16 L和TXNRD 1。(C)2014爱思唯尔有限公司版权所有。
The role in cancer chemoprevention of proteins present in non-digestible fraction of common bean (Phaseolus vulgaris L.) is unknown. The aim was to examine the effect of peptide extracted from two cultivars of the common bean Azufrado Higuera (AH-PE) and Bayo Madero (BM-PE) on a gene expression using human colon cancer cells through a microarray analysis. HCT116 and RKO colon cancer cells were treated with 0.5 mg/mL AH-PE and BM-PE for 24 h; RNA was extracted and the gene expression was analyzed using an Agilent Low Input Quick Amp Labeling assay. In the HCT116 human cells, 511 and 964 genes were affected by the AH-PE and BM-PE treatments, respectively, with 405 genes found to be commonly affected by peptide extracts. In the RKO human cells, 45 and 32 genes were affected significantly by the AH-PE and BM-PE treatments, respectively, with 19 genes common for both extracts. Bioinformatic analysis using Database for Annotation Validation and Integrated Discovery showed that the primary genes affected were involved with oxidation reduction, response to stimulus, protein phosphatase, MAPK signaling, selenium binding and response to wounding. Ingenuity Pathway Analysis showed that in the HCT116 cell line, NRF-2 related antioxidant enzymes were up-regulated while glutathione redox-related enzymes were down-regulated in the RKO cell line, potentially causing reactive oxygen species (ROS) imbalance. Validation of genes by RT-PCR showed good agreement for the genes with a potential role in cancer proliferation and apoptosis (PDE4B and OSGIN1, and KRT19 and EEF1A2). This study discovered, for the first time, 8 differentially expressed genes to be similar for both human colon cancer cell lines triggered by both bean cultivars, namely C11orf31, C9orf169, EMP1, GEM, PLIN2, SUN3, TRIM16L and TXNRD1. (C) 2014 Elsevier Ltd. All rights reserved.