Nair Sujit, Xu Changjiang, Shen Guoxiang, Hebbar Vidya, Gopalakrishnan Avantika, Hu Rong, Jain Mohit Raja, Lin Wen, Keum Young-Sam, Liew Celine, Chan Jefferson Y, Kong Ah-Ng Tony
Graduate Program in Pharmaceutical Sciences, Department of Pharmaceutics, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, Piscataway, New Jersey 08854, USA.
Pharm Res. 2006 Nov;23(11):2621-37. doi: 10.1007/s11095-006-9099-x. Epub 2006 Sep 13.
The objective of this study was to investigate the pharmacogenomics and the spatial regulation of global gene expression profiles elicited by cancer chemopreventive agent butylated hydroxyanisole (BHA) in mouse small intestine and liver as well as to identify BHA-modulated nuclear factor-E2-related factor 2 (Nrf2)-dependent genes.
C57BL/6J (+/+; wildtype) and C57BL/6J/Nrf2(-/-; knockout) mice were administered a single 200 mg/kg oral dose of BHA or only vehicle. Both small intestine and liver were collected at 3 h after treatment and total RNA was extracted. Gene expression profiles were analyzed using 45,000 Affymetrix mouse genome 430 2.0 array and GeneSpring 7.2 software. Microarray results were validated by quantitative real-time reverse transcription-PCR analyses.
Clusters of genes that were either induced or suppressed more than two fold by BHA treatment compared with vehicle in C57BL/6J/Nrf2(-/-; knockout) and C57BL/6J Nrf2 (+/+; wildtype) mice genotypes were identified. Amongst these, in small intestine and liver, 1,490 and 493 genes respectively were identified as Nrf2-dependent and upregulated, and 1,090 and 824 genes respectively as Nrf2-dependent and downregulated. Based on their biological functions, these genes can be categorized into ubiquitination/proteolysis, apoptosis/cell cycle, electron transport, detoxification, cell growth/differentiation, transcription factors/interacting partners, kinases and phosphatases, transport, biosynthesis/metabolism, RNA/protein processing and nuclear assembly, and DNA replication genes. Phase II detoxification/antioxidant genes as well as novel molecular target genes, including putative interacting partners of Nrf2 such as nuclear corepressors and coactivators, were also identified as Nrf2-dependent genes.
The identification of BHA-regulated and Nrf2-dependent genes not only provides potential novel insights into the gestalt biological effects of BHA on the pharmacogenomics and spatial regulation of global gene expression profiles in cancer chemoprevention, but also points to the pivotal role of Nrf2 in these biological processes.
本研究的目的是调查癌症化学预防剂丁基羟基茴香醚（BHA）在小鼠小肠和肝脏中引发的药物基因组学以及全球基因表达谱的空间调控，并鉴定BHA调节的核因子E2相关因子2（Nrf2）依赖性基因。
给C57BL/6J（+/+；野生型）和C57BL/6J/Nrf2（-/-；敲除）小鼠口服单次剂量200mg/kg的BHA或仅给予溶剂。在治疗后3小时收集小肠和肝脏，并提取总RNA。使用45,000个Affymetrix小鼠基因组430 2.0阵列和GeneSpring 7.2软件分析基因表达谱。通过定量实时逆转录-PCR分析验证微阵列结果。
在C57BL/6J/Nrf2（-/-；敲除）和C57BL/6J Nrf2（+/+；野生型）小鼠基因型中，鉴定出与溶剂相比经BHA处理诱导或抑制超过两倍的基因簇。其中，在小肠和肝脏中，分别有1490个和493个基因被鉴定为Nrf2依赖性且上调，分别有1090个和824个基因被鉴定为Nrf2依赖性且下调。根据其生物学功能，这些基因可分为泛素化/蛋白水解、凋亡/细胞周期、电子传递、解毒、细胞生长/分化、转录因子/相互作用伴侣、激酶和磷酸酶、转运、生物合成/代谢、RNA/蛋白质加工和核组装以及DNA复制基因。II期解毒/抗氧化基因以及新的分子靶基因，包括Nrf2的推定相互作用伴侣如核共抑制因子和共激活因子，也被鉴定为Nrf2依赖性基因。
BHA调节的和Nrf2依赖性基因的鉴定不仅为BHA在癌症化学预防中对药物基因组学和全球基因表达谱的空间调控的整体生物学效应提供了潜在的新见解，也指出了Nrf2在这些生物学过程中的关键作用。
