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cytochrome b5 reductase 2 suppresses tumor formation in nasopharyngeal carcinoma by attenuating angiogenesis

Chinese Journal of Cancer 2015年第10期34卷 459-467页

作者： Huixin Ming Ying Lan Feng He Xue Xiao Xiaoying Zhou Zhe Zhang Ping Li Guangwu Huang Department of Pathology First Affiliated Hospital of Guangxi Medical University Department of Otolaryngology-Head and Neck Surgery First Affiliated Hospital of Guangxi Medical University Medical Research Centre Guangxi Medical University

Background:cytochrome b5 reductase 2(CYB5R2) is a potential tumor suppressor that inhibits cell proliferation and motility in nasopharyngeal carcinoma(NPC).Inactivation of CYB5R2 is associated with lymph node metastasis in NPC.This study aimed to explore the mechanisms contributing to the anti-neoplastic effects of CYB5R2.Methods:Polymerase chain reaction(PCR) assays were used to analyze the transcription of 84 genes known to be involved in representative cancer pathways in the NPC cell line HONE1.NPC cell lines CNE2 and HONE1 were transiently transfected with CYB5R2,and data was validated by real-time PCR.A chick chorioallantoic membrane(CAM)embryo model was implanted with CYB5R2-expressing CNE2 and HONE1 cells to evaluate the effect of CYB5R2 on angiogenesis.An immunohistochemical assay of the CAM model was used to analyze the protein expression of vascular endothelial growth factor(VEGF).Results:In CYB5R2-transfected NPC cells,PCR assays revealed up-regulated mRNA levels of Fas cell surface death receptor(FAS),FBJ murine osteosarcoma viral oncogene homolog(FOS),phosphoinositide-3-kinase regulatory subunit 1(PIK3R1),integrin beta 3(ITGB3),metastasis suppressor 1(MTSSl),interferon beta 1(IFNB1),and cyclin-dependent kinase inhibitor 2A(CDKN2A) and down-regulated levels of integrin beta 5(ITGB5),insulin-like growth factor 1(IGF1),TEK tyrosine kinase(TEK),transforming growth factor beta receptor 1(TGFBRl),and VEGF.The angiogenesis in the CAM model implanted with CYB5R2-transfected NPC cells was inhibited.Down-regulation of VEGF by CYB5R2 in NPC cells was confirmed by immunohistochemical staining in the CAM model.Conclusion:CYB5R2 up-regulates the expression of genes that negatively modulate angiogenesis in NPC cells and down-regulates the expression of VEGF to reduce angiogenesis,thereby suppressing tumor formation.

关键词： cytochrome b5 reductase 2, Nasopharyngeal carcinoma, Chick embryo model, Angiogenesis

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An update on the role of intestinal cytochrome P450 enzymes in drug disposition

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Acta Pharmaceutica Sinica B 2016年第5期6卷 374-383页

作者： Fang Xie Xinxin Ding Qing-Yu Zhang Bioimaging Glaxo Smith KlineKing of Prussia College of Nanoscale Science SUNY Polytechnic Institute Wadsworth Center New York State Department of Health

Oral administration is the most commonly used route for drug treatment.Intestinal cytochrome P450(CYP)-mediated metabolism can eliminate a large proportion of some orally administered drugs before they reach systemic circulation,while leaving the passage of other drugs unimpeded.A better understanding of the ability of intestinal P450 enzymes to metabolize various clinical drugs in both humans and preclinical animal species,including the identification of the CYP enzymes expressed,their regulation,and the relative importance of intestinal metabolism compared to hepatic metabolism,is important for improving bioavailability of current drugs and new drugs in development.Here,we briefly review the expression of drug-metabolizing P450 enzymes in the small intestine of humans and several preclinical animal species,and provide an update of the various factors or events that regulate intestinal P450 expression,including a cross talk between the liver and the intestine.We further compare various clinical and preclinical approaches for assessing the impact of intestinal drug metabolism on bioavailability,and discuss the utility of the intestinal epithelium–specific NADPH-cytochrome P450 reductasenull(IECN) mouse as a useful model for studying in vivo roles of intestinal P450 in the disposition of orally administered drugs.

关键词： cytochrome P450 Intestine Bioavailability Drug disposition Drug metabolism

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Berberine Induces Cell Apoptosis through cytochrome C/Apoptotic Protease-Activating Factor 1/Caspase-3 and Apoptosis Inducing Factor Pathway in Mouse Insulinoma Cells

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Chinese Journal of Integrative Medicine 2019年第11期25卷 853-860页

作者： FANG Xin MIAO Xiao-liang LIU Jun-li ZHANG Dong-wei WANG Min ZHAO Dan-dan MU Qian-qian YU Na MO Fang-fang YIN Hong-ping GAO Si-hua School of Basic Medicine Beijing University of Chinese MedicineBeijing (100029)China Fraser Laboratories for Diabetes Research Department of MedicineRI-McGill University Health CentreMontrealQuebec (H4A3J1)Canada School of Life Science and Tech no logy China Pharmaceutical Un iversityNanjing (210009)China Diabetes Research Center Beijing University of Chinese MedicineBeijing (100029)China

Objective:To investigate apoptotic effects of berberine,a significant alkaloids component existing in Rhizoma coptidis,and its possible acting mechanism in insulinoma cells.Methods:Different concentrations of berberine were used to treat mouse insulinoma(MIN6)cells for various period of time.The viability and apoptosis of the cells were analyzed using methylthiazolyldiphenvl-tetrazolium bromide assay,flow cytometry and enzyme-linked immuno sorbent assay.Changes in the relating pro-and anti-apoptosis proteins were detected by western-blotting.Results:The half-maximal inhibitory concentration(IC50)of berberine was 5.7μmol/L on MIN6 cells viability for 16 h.Berberine caused a 20%reduction(Pcytochrome C transforming from the mitochondria to the cytoplasm.Apoptotic protease-activating factor 1(Apaf-1)was subsequently activated after cytochrome C release.Furthermore,caspase-3 and poly adenosine diphosphate-ribose polymerase were also activated to trigger apoptosis cascade.Conclusion:High concentration(5 and 10μmol/L)of berberine could induce the apoptosis of MIN6 cells through cytochrome C/Apaf-1/caspase-3 and apoptosis inducing factor(AIF)pathway.

关键词： Chinese medicine berberine apoptosis mouse insulinoma cells cytochrome C/Apaf-1/caspase-3 apoptosis inducing factor pathway

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Identification of the 2-tridecanone cis-acting element in the promoter of cytochrome P450 CYP6B7 in Helicoverpa armigera

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Insect Science 2018年第6期25卷 959-968页

作者： Li Xu Dong-Zhi Li Yuan-Yuan Luo Jian-Ying Qin Li-Hong Qiu College of Science China Agricultural UniversityBeijingChina

The expression level of cytochrome P450 genes in insects can be induced by plant allelochemicals,which is important for insects to adapt to host plants.cytochrome P450CYP6B 7has been reported to be involved in pyethroid insecticide resistance in Heli- coverpa armigera,and its transcription level was induced by some inducers.Currently,the regulatory mechanism of the induced expression of CYP6B7remains unknown,although it is very important for understanding the detoxification mechanism to allelochemicals in host plants.The objective of the present study was to investigate the eis-acting ele- ment in the promoter of CYP6B7 mediating the inducible up-regulation of CYP6B7in H.armigera by 2-tridecanone.The promoter region of CYP6B7was cloned by genome walking technique and analyzed by transient transfeetion assay.Progressive 5'deletion of the promoter region of CYP6B7revealed that the relative luciferase activity of construct -320/+232could be significantly induced by 2-trideeanone.Further stepwise deletion between -320 and -238 bp found that construct -292/+232 could also be significantly induced by 2-tridecanone,but the adjacent construct -256/+232could not,suggesting the essential role of the sequence between -292 and --257 bp for 2-tridecanone induction. Nucleotide mutations between -292 and -281 bp had no influence on the induction ef- fect by 2-tridecanone,but nucleotide mutations between -280 and -257 bp significantly decreased the induction effect.These results demonstrated that the cis-acting element for 2-trideeanone induction was between -280 and -257 bp in the promoter of CYP6B7.

关键词： cis-acting element cytochrome P450 CYP6B7 Helicoverpa armigera induction 2-tridecanone

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Identification and analysis of a processed cytochrome P450 pseudogene of the disease vector Aedes aegypti

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Asian Pacific Journal of Tropical Medicine 2016年第10期9卷 951-956页

作者： Fatma M.A.El-garj Mustafa F.F.Wajidi Silas W.Avicor Molecular Entomology Research Group School of Distance EducationUniversiti Sains Malaysia

Objective: To clone cytochrome P450 from Aedes aegypti(Ae. aegypti) and determine the characteristics using bioinformatics tools. Methods: cytochrome P450 of Ae. aegypti was amplified using polymerase chain reaction, cloned and sequenced. Evolutionary relationship of the sequence was inferred and bioinformatics tools were used to predict subcellular localisation, signal peptide, transmembrane helix, phosphorylation, O-glycosylation, secondary and tertiary structures of the deduced protein. Results: Polymerase chain reaction rather amplified a cytochrome P450 pseudogene which was named CYP4H44P(Gen Bank accession number KF779932). The pseudogene has 1537 nucleotides and an open reading frame of 335 amino acids containing cytochrome P450 motifs except the Wxxx R motif. It is highly homologous to CYP4H28 and CYP4H28v2. Phylogenetic analysis and evolutionary divergence showed strong clustering with CYP4H28 alleles and least divergence from the alleles respectively. The deduced protein was predicted to be found in the cytoplasm and likely to be phosphorylated but devoid of signal peptide, transmembrane helix and O-glycosylated sites. The secondary and tertiary structures were also generated. Conclusions: A cytochrome P450 pseudogene, CYP4H44 P was cloned from Ae. aegypti. The pseudogene is homologous with CYP4H28 alleles and seems to have recently diverged from this group. Isolating this pseudogene is an important step for evaluating its biological role in the mosquito and for the evolutionary analysis of Ae. aegypti CYPs.

关键词： Aedes aegypti Clone cytochrome P450 pseudogene CYP4H44P Bioinformatics

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Role of cytochrome P450 polymorphisms and functions in development of ulcerative colitis

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World Journal of Gastroenterology 2019年第23期25卷 2846-2862页

作者： Alaattin Sen Holger Stark Department of Molecular Biology and Genetics Faculty of Life and Natural SciencesAbdullah Gul UniversityKayseri 38080Turkey Biology Department Faculty of Arts and SciencesPamukkale UniversityDenizli 20070Turkey Institute of Pharmaceutical and Medicinal Chemistry Heinrich Heine University DüsseldorfDuesseldorf 40225Germany

cytochromes P450 s(CYPs) are terminal enzymes in CYP dependent monooxygenases, which constitute a superfamily of enzymes catalysing the metabolism of both endogenous and exogenous substances. One of their main tasks is to facilitate the excretion of these substances and eliminate their toxicities in most phase 1 reactions. Endogenous substrates of CYPs include steroids, bile acids, eicosanoids, cholesterol, vitamin D and neurotransmitters. About 80% of currently used drugs and environmental chemicals comprise exogenous substrates for CYPs. Genetic polymorphisms of CYPs may affect the enzyme functions and have been reported to be associated with various diseases and adverse drug reactions among different populations. In this review, we discuss the role of some critical CYP isoforms(CYP1 A1, CYP2 D6, CYP2 J2, CYP2 R1,CYP3 A5, CYP3 A7, CYP4 F3, CYP24 A1, CYP26 B1 and CYP27 B1) in the pathogenesis or aetiology of ulcerative colitis concerning gene polymorphisms. In addition, their significance in metabolism concerning ulcerative colitis in patients is also discussed showing a clear underestimation in genetic studies performed so far.

关键词： cytochrome P450 Polymorphism Function Ulcerative colitis Aetiology

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Effect of urea on synchronous fluorescence spectra and electrochemical behaviour of cytochrome

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Science China Chemistry 1996年第2期39卷 211-218页

作者：侴菊陆天虹吴越 Laboratory of Electroanalytical Chemistry Changchun Institute of Applied Chemistry Chinese Academy of Sciences Changchun 130022 China

The changes of the synchronous fluorescence spectra and the electrochemical behaviour of cytochrome c with the urea concentration are studied. It has been found that with the increase of urea concentration, there occur sequentially the deaggregation of cytochrome c molecules, the increase of exposure extent of the heme group to the solvent, the disruption of Fe-S bond of the heme group and the change in the electrochemical behaviour of cytochrome c. It is suggested that the reason why the electrochemical reaction of cytochrome c is irreversible is that cytochrome c molecules exist in the concentrated solution as oligomers which are electrochemically inactive.

关键词： cytochrome c urea synchronous fluorescence spectroscopy.

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Compound heterozygous variants in POR gene identified by whole-exome sequencing in a Chinese pedigree with cytochrome P450 oxidoreductase deficiency

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Pediatric Investigation 2018年第2期2卷 90-95页

作者： Chanjuan Hao Jun Guo Ruolan Guo Zhan Qi Wei Li Xin Ni Beijing Key Laboratory for Genetics of Birth Defects MOE Key Laboratory of Major Diseases in Children Center for Medical Genetics Beijing Pediatric Research Institute Beijing Children's Hospital Capital Medical UniversityNational Center for Children's HealthBeijing China Beijing Pediatric Research Institute Department of Otolaryngology Head and Neck Surgery Beijing Children's Hospital Capital Medical UniversityNational Center for Children's HealthBeijing China

Importance:cytochrome P450 oxidoreductase deficiency (PORD) is a rare disease exhibiting a variety of clinical manifestations.This condition specifically leads to disordered steroidogenesis,which can affect the development of the reproductive system,skeleton,and other parts of the body.The severe form of PORD is difficult to differentiate with Antley-Bixler syndrome (ABS).The genetic characters and clinical evaluation of PORD are still unclear in China.Objective:To perform an exome analysis and identify the pathogenic cause in order to assist clinicians to obtain a proper evaluation on the genetic condition.Methods:The proband underwent detailed physical evaluations.DNA of the proband and his parents was isolated and whole-exome sequencing (WES) was performed.Variants were analyzed and evaluation according to the ACMG guideline.Results:A 1-year-old Chinese boy with midface hypoplasia,choanal stenosis,multiple joint contractures,micropenis and right cryptorchidism was misdiagnosed with Crouzon syndrome.By trio-whole-exome sequencing,we identified an unreported compound heterozygous mutation (c.667C＞T,p.R223* and c.1370G＞A,p.R457H) in POR in the proband.This mutation was inherited from healthy heterozygous parents,supporting the diagnosis of PORD,which was further confirmed by biochemical characteristics.Interpretation:We have identified a pathogenic variant with an unreported compound heterozygous POR mutation,which expands the clinical and genetic spectra of PORD and emphasizes the usefulness of WES for genetic diagnosis.

关键词： Compound heterozygous mutation cytochrome P450 oxidoreductase Disordered steroidogenesis POR deficiency Whole-exome sequencing

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Characteristics and roles of cytochrome b5 in cytochrome P450-mediated oxidative reactions in Locusta migratoria

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Journal of Integrative Agriculture 2020年第6期19卷 1512-1521页

作者： LIU Jiao ZHANG Xue-yao WU Hai-hua MA Wen ZHU Wen-ya Kun-Yan ZHU MA En-bo ZHANG Jian-zhen Institute of Applied Biology Shanxi UniversityTaiyuan 030006P.R.China College of Life Science Shanxi UniversityTaiyuan 030006P.R.China Modern Research Center for Traditional Chinese Medicine Shanxi UniversityTaiyuan 030006P.R.China Institute of Plant Protection Shanxi Academy of Agricultural SciencesTaiyuan 030031P.R.China Department of Entomology Kansas State UniversityManhattanKS 66506USA

cytochrome b5(Cyt-b5)is a small heme protein and known to be involved in a wide range of biochemical transformations,in eluding cytochrome P450 monooxyge nase(CYP)-mediated metabolism of endoge nous and exogenous compo un ds.Studies on Cyt-b5 are more con centrated in mammals,but are relatively rare in in sects.The characteristics and functi on of Cyt-b5 from Locusta migratoria have not been described yet.We sequeneed the full-length cDNA sequenee of Cyt-b5 from L.migratoria(LmCyt-b5)by reverse transcription-PCR(RT-PCR)based on locust transcriptome database.The phylogenetic analysis showed that LmCyt-b5 was closely related to the Cyt-b5 from Blattodea.LmCyt-b5 was highly expressed in ovary,Malpighian tubules,midgut,gastric caeca,and fat bodies.Silencing of LmCyt-b5 had no effect on the susceptibility of L.migratoria to four different insecticides.Suppression of LmCyt-b5 or silencing of both LmCyt-b5 and LmCPR did not significantly change the total CYP activity toward the substrate 7-ethoxycoumarin(7-EC).However,coexpression of LmCYP6FD1 with LmCPR and LmCyt-b5 together in Sf9 cells by using Bac-to-Bac baculovirus expression system significantly increased the catalytic activity of LmCYP6FD1 toward 7-EC as compared with the coexpression of L.mCYP6FD1 with cytochrome P450 reductase(LmCPR)or LmCyt-b5 separately.These results suggest that LmCyt-b5 plays an important role in the catalytic reaction of LmCYP6FD1 toward 7-EC in our in vitro experiments.Further study is needed to clarify the role of LmCyt-b5 in CYP-mediated catalytic reactions in L.migratoria.

关键词： cytochrome b5 cytochrome P450 cytochrome P450 reductase Locusta migratoria RNA interference

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NMR studies of the effects of axial ligand substitution by imidazole on the conformation of cytochrome c

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Science China Chemistry 1995年第6期38卷 649-657页

作者：邵伟平姚亦明唐文霞 State Key Laboratory of Coordination Chemistry Nanjing University Nanjing 210093 China

A series of 1D and 2D 1H NMR studies are presented, which aim at assigning the hyperfine shifted resonances of the heme and the spin systems of more than 20 amino acid side chains, and characterizing the heme and ligand environment of the low spin imidazole complex of cytochrome c. Some details of the structure in the heme pocket of the cytochrome c-imidazole complex are shownqualitatively to have changed relatively to the native oxidized protein. The orientation of the coordinated imidazole is also determined based on the interpretation of a number of NOEs.

关键词： imidazole complex of cytochrome c NMR axial ligand conformation.

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