维生素A和D的联合诱导在乳腺癌症细胞中具有协同作用,并改变内质网应激、未折叠蛋白和雌激素信号传导经典途径中的基因表达

IF 1.3 Q4 FOOD SCIENCE & TECHNOLOGY
Pinal N. Kanabar, Nina S. Los, T. Lawal, S. Patel, Nisikant A. Raut, M. Maienschein-Cline, Z. Arbieva, G. Mahady
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引用次数: 1

摘要

简介:过去30年的大量研究表明,功能性食品中存在的生物活性化合物,包括维生素、矿物质和植物化学物质,可以降低癌症的风险。例如,水果、蔬菜和乳制品中的维生素A和D衍生物可以减少乳腺癌、膀胱癌、头颈癌、肺癌、前列腺癌和皮肤癌的生长。然而,这些联合维生素对癌症的作用以前还没有报道过。目的:研究维生素A(全反式维甲酸;ATRA)以及维生素D2和D3在乳腺上皮癌症细胞系T47D:A18、MCF-7和SK-BR-3中的联合活性,并使用RNA-seq对MCF-7细胞进行全基因组分析,或1、5和10μg/ml的组合。CellTiter-Glo®2.0测定法、Caspase-Glo®3/7、Caspase®8和ApoTox-Glo™ Triplex测定法测定细胞活力和细胞凋亡。用CYTO-ID®自噬检测试剂盒2.0测量治疗对MCF-7细胞自噬的影响。使用mRNA-seq和qPCR评估整个转录组分析。结果:分别,ATRA、D2和D3均降低了所有测试的乳腺癌症细胞系的生存能力,中位抑制浓度(IC50)在2.1至31.7μg/ml之间。然而,当用ATRA+D2+D3的组合处理癌症乳腺细胞时,IC50降低,表明协同作用。在MCF-7细胞中,5-氟脲嘧啶(5-FLU)的IC50为1.37µg/ml,而维生素A和D组合的IC50则为1.5µg/ml,表明该组合的有效性约为5-FLU的90%。ATRA+D2+D3处理MCF-7细胞增强了胱天蛋白酶3/7的活性,以及Bax、BAD、PTEN和p53(凋亡典型途径)的表达,以及诱导的自噬。经处理的MCF-7细胞的全基因组分析显示,自噬、内质网应激和未折叠蛋白反应细胞凋亡典型途径中的基因表达显著上调。此外,用ATRA+D2+D3处理的MCF-7细胞在雌激素介导的S期进入和雌激素信号传导经典途径中表现出显著的基因表达下调,这表明其具有抗雌激素作用。结论:维生素A和维生素D联合用药对癌症细胞具有协同作用,可诱导细胞凋亡和自噬。转录谱显示基因表达模式的显著改变和多种癌症信号通路的上调支持了维生素A和D联合使用比单独使用维生素更有效的治疗假设。关键词:全反式维甲酸(ATRA),维生素D2,维生素D3,乳腺癌症细胞,凋亡,雌激素受体
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Combinations of vitamin A and D induced are synergistic in breast cancer cells and alter gene expression in the endoplasmic reticulum stress, unfolded protein and estrogen signaling canonical pathways
Introduction: Numerous studies over the past 30 years have shown that bioactive compounds present in functional foods, including vitamins, minerals and phytochemicals reduce cancer risk. For example, vitamins A and D derivatives found in fruits, vegetables, and dairy products, reduce the growth of breast, bladder, head, neck, lung, prostate, and skin cancers. However, the effects of these combined vitamins have not been previously reported for breast cancer.Aims: To investigate the activities of vitamin A (all-trans-retinoic acid; ATRA), as well as vitamins D2 and D3 in combination in the breast epithelial cancer cell lines T47D:A18, MCF-7, and SK-BR-3 and perform whole genome analysis in MCF-7 cells using RNA-seq.Methods: Breast cancer cells were cultured in appropriate media and treated with ATRA, D2 or D3 alone in concentrations from 1-10 μg/ml, or in combination at 1, 5, and 10 μg/ml. The CellTiter-Glo® 2.0 assay, Caspase-Glo®3/7, Caspase®8, and ApoTox-Glo™ Triplex assays measured cell viability and apoptosis. The effect of treatment on autophagy in MCF-7 cells was measured with a CYTO-ID® Autophagy Detection Kit 2.0. The whole transcriptome analysis was assessed using mRNA-seq and qPCR.Results: Separately, ATRA, D2, and D3 all reduced the viability of all breast cancer cell lines tested, with median inhibitory concentrations (IC50) between 2.1 to 31.7 μg/ml. However, when breast cancer cells were treated with combinations of ATRA+D2+D3, the IC50 was reduced indicating synergism. In MCF-7 cells, 5-flurouracil (5-FLU) had an IC50 of 1.37 µg/ml, while the vitamin A and D combination had an IC50 of 1.5 µg/ml, indicating the combination was ~90% as effective as 5-FLU. Treatment of MCF-7 cells with ATRA+D2+D3 enhanced caspase 3/7 activity, as well as the expression of Bax, BAD, PTEN and p53 (apoptosis canonical pathway), as well as induced autophagy. Whole genome analysis of treated MCF-7 cells showed a significant upregulation in gene expression in the autophagy, endoplasmic reticulum stress and the unfolded protein response apoptosis canonical pathways. Furthermore, MCF-7 cells treated with ATRA+D2+D3 demonstrated significant downregulation of gene expression in estrogen-mediated S phase entry and estrogen signaling canonical pathways suggesting antiestrogenic effects. Conclusions: Vitamins A and D combinations had synergistic effects in breast cancer cells and induced both apoptosis and autophagy. Transcriptional profiling showed significant alterations in gene expression patterns and upregulation of multiple cancer signaling pathways supporting the hypothesis that combining vitamins A and D is a more effective treatment than either vitamin alone. Keywords: all-trans-retinoic acid (ATRA), Vitamin D2, Vitamin D3, breast cancer cells, apoptosis, estrogen receptors
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来源期刊
Functional Foods in Health and Disease
Functional Foods in Health and Disease FOOD SCIENCE & TECHNOLOGY-
CiteScore
2.20
自引率
20.00%
发文量
47
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