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Quantitative Evaluation of aldo-keto reductase Expression in Hepatocellular Carcinoma (HCC) Cell Lines

Genomics, Proteomics & Bioinformatics 2013年第4期11卷 230-240页

作者： Lei Yang Ju Zhang Shenyan Zhang Weiwei Dong Xiaomin Lou Siqi Liu CAS Key Laboratory of Genome Sciences and Information Beijing Institute of Genomics Chinese Academy of Sciences Department of Oncology General Hospital of PLA

The involvement of aldo-keto reductases （AKRs） in tumorigenesis is widely reported, but their roles in the pathological process are not generally recognized due to inconsistent measure- ments of their expression. To overcome this problem, we simultaneously employed real-time PCR to examine gene expression and multiple reaction monitoring （MRM） of mass spectrometry （MS） to examine the protein expression of AKRs in five different hepatic cell lines. These include one rela- tively normal hepatic cell line, L-02, and four hepatocellular carcinoma （HCC） cell lines, HepG2, HUH7, BEL7402 and SMMC7721. The results of real-time PCR showed that expression of genes encoding the AKR1C family members rather than AKR1A and AKR1B was associated with tumor, and most of genes encoding AKRs were highly expressed in HUH7. Similar observations were obtained through MRM. Different from HUH7, the protein abundance of AKR1A and AKR1B was relatively consistent among the other four hepatic cell lines, while protein expression of AKR1C varied significantly compared to L-02. Therefore, we conclude that the abundant distri- bution of AKR 1C proteins is likely to be associated with liver tumorigenesis, and the AKR expres- sion status in HuH7 is completely different from other liver cancer cell lines. This study, for the first time, provided both overall and quantitative information regarding the expression of AKRs at both mRNA and protein levels in hepatic cell lines. Our observations put the previous use of AKRs as a biomarker into question since it is only consistent with our data from HUH7. Furthermore, the data presented herein demonstrated that quantitative evaluation and comparisons within a protein fam- ily at both mRNA and protein levels were feasible using current techniques.

关键词： aldo-keto reductase HCC Quantitative analysis Real-time PCR Multiple reaction monitoring

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Expression Analysis of aldo-keto reductase 1 (AKR1) in Foxtail Millet (Setaria italica L.) Subjected to Abiotic Stresses

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American Journal of Plant Sciences 2016年第3期7卷 500-509页

作者： Tanguturi Venkata Kirankumar Kalaiahgari Venkata Madhusudhan Ambekar Nareshkumar Kurnool Kiranmai Uppala Lokesh Boya Venkatesh Chinta Sudhakar Plant Molecular Biology Laboratory Department of Botany Sri Krishnadevaraya University Anantapuramu India

Foxtail millet (Setaria italica L.) is a drought-tolerant millet crop of arid and semi-arid regions. aldo-keto reductases (AKRs) are significant part of plant defence mechanism, having an ability to confer multiple stress tolerance. In this study, AKR1 gene expression was studied in roots and leaves of foxtail millet subjected to different regimes of PEG- and NaCl-stress for seven days. The quantitative Real-time PCR expression analysis in both root and leaves showed upregulation of AKR1 gene during PEG and salt stress. A close correlation exits between expression of AKR1 gene and the rate of lipid peroxidation along with the retardation of growth. Tissue-specific differences were found in the AKR1 gene expression to the stress intensities studied. The reduction in root and shoot growth under both stress conditions were dependent on stress severity. The level of lipid peroxidation as indicated by MDA formation was significantly increased in roots and leaves along with increased stress levels. Finally, these findings support the early responsive nature of AKR1 gene and seem to be associated at least in part with its ability to contribute in antioxidant defence related pathways which could provide a better protection against oxidative stress under stress conditions.

关键词： aldo-keto reductase Lipid Peroxidation Reactive Carbonyls Cellular Homeostasis Plant Abiotic Stress Response

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Genetic analysis and functional identification of tetrahydrobiopterin synthesis-related silkworm mutants

Genetic analysis and functional identification of tetrahydro...

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中国蚕学会第十届青年学术研讨会

作者： MENG Yan SHU Rui MIN Qin QIN Hongwei CAI Yangyang GAO Junshan School of life sciences Anhui Agricultural University

Tetrahydrobiopterin(BH4) is an essential cofactor for aromatic amino acid hydroxylases and nitric oxide synthase(NOS) so that BH4 controls the levels of monoamine neurotransmitters of dopamine and serotonin and signaling molecule of NO in vivo,playing a key role in many biological ***4 deficiency is associated with numerous metabolic syndromes and neuropsychological *** recent years’ researches found that the de novo synthesis pathway of BH4 is similar between insects and *** the silkworm Bombyx mori,there are several genetic mutants potentially involving in BH4 synthesis pathway,such as lem and lem lethal allele(lem),al and i-lem(inhibitor of lem),which are good models to comprehensively study the regulation mechanism of BH4 synthesis in *** have made it clearly that genetic mutation of sepiapterin reductase(BmSPR),a key enzyme acting on the BH4 synthesis pathway,is responsible for the silkworm lem and lem mutants due to the down-regulation of ***,physical lethality of leml larvae is still *** i-lem inhibits lem phenotype at the molecular level is also *** recent reasearches found that a mutation in the aldo-keto reductase 2 gene(BmAkr) is causative in the i-lem mutant by functioning at the alternative BH4 synthesis *** addition,a nitric oxide synthase gene of ***(BmNOS1) was *** transcription and coupling of BmNOS1 because of BH4 deficiency led to the reduction of NO and increase of superoxide anion,which might contribute to the cell damage and death in the lem larvae.

关键词： Bombyx mori lem and i-lem mutants tetrahydrobiopterin sepiapterin reductase aldo-keto reductase nitric oxide synthase

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