Cloning of TaCYP707A1 Gene that Encodes ABA 8′-Hydroxylase in Common Wheat (Triticum aestivum L.)

The plant hormone abscisic acid （ABA） regulates many important physiological and developmental processes in plants. The objective of this study was to clone the ABA 8′-hydroxylase gene in common wheat. In the present study, we used the eDNA sequence of barley HvCYP707A1 gene （GenBank accession no. AB239299） as a probe for BLAST search against the common wheat （Triticum aestivum L.） EST database in GenBank. All wheat ESTs sharing high similarity with the reference gene were subjected to contig assembly. Primers were designed based on the constructed contigs to clone the wheat CYP707A1 gene, designated as TaCYP707A1. The genomic DNA sequence of TaCYPTO7A1 gene comprised five exons and four introns, with a size of 2225 bp. The corresponding cDNA sequence of TaCYP707A1 was 1737 bp, containing an open reading frame （ORF） of 1431 bp, a 42-bp 5′-untranslated region （UTR） and a 264-bp 3′UTR, with 94.9% of identical sequences to HvCYP707A1 gene （AB239299）. The neighbor joining tree indicated that the deduced amino acid sequences of TaCYP707A1 gene was highly similar to those of barley and rice. The TaCYP707A1 gene was located on chromosome 6BL using a set of Chinese Spring nullisomic-tetrasomic lines and ditelosomic line 6BS. These results will be of high importance in understanding of molecular mechanism of ABA catabolism.

Polymorphisms in CYP2R1 Gene Associated with Serum Vitamin D Levels and Status in a Chinese Rural Population

Vitamin D, a fat-soluble vitamin and endocrine horm one, and it impacts various bone and extra-bone health, such as osteoporosis, diabetes, and cancer. The main circulating form of vitamin D is 25-hydroxyvitamin D [25(OH)D] and it is a useful clinical biomarker of vitamin D status. The Institute of Medicine (IOM) defines as vitamin D deficiency (VDD) when serum 25(OH)D concentration is less than 20 ng/mL⑴.Worldwide, VDD is recognized as a severe public health problem. In 2007, Holick estimated that globally over one billion people suffered from VDD or vitamin D insufficiency (VDI). In China, it has bee n reported that the prevale nee of VDD ranged from 38.8% to 91.2% in different regions.

Compound heterozygous mutations in CYP1B1 gene leads to severe primary congenital glaucoma phenotype

AIM: To identify the novel mutation alleles in the CYP1B1 gene of primary congenital glaucoma(PCG) patients at Shandong Province of China, and investigate their correlation with glaucomatous features.METHODS: The DNA from the peripheral blood of 13 congenital glaucoma patients and 50 ethnically matched healthy controls from the affiliated hospital of Qingdao University were extracted. The coding region of the CYP1B1 gene was amplified by PCR and direct DNA sequencing was performed. Disease causing-variants were analyzed by comparing the sequences and the structures of wild type and mutant CYP1B1 proteins by PyMOL software.RESULTS: Two missense mutations, including A330 F caused by c.988 G>T&c.989 C>T, and R390H caused by c.1169 G>A, were identified in one of the 13 PCG patients analyzed in our study. A330F mutation was observed to be novel in the Chinese Han population, which dramatically altered the protein structure of CYP1B1 gene, including the changes in the ligand-binding pocket. Furthermore, R390H mutation caused the changes in heme-protein binding site of this gene. In addition, the clinical phenotype displayed by PCG patient with these mutations was more pronounced than other PCG patients without these mutations. Multiple surgeries and combined drug treatment were not effective in reducing the elevated intraocular pressure in this patient.CONCLUSION: A novel A330F mutation is identified in the CYP1B1 gene of Chinese PCG patient. Moreover, in combination with other mutation R390H, this PCG patient shows significant difference in the CYP1B1 protein structure, which may specifically contribute to severe glaucomatous phenotype.

Cigarette smoking, dietary habits and genetic polymorphisms in GSTT1, GSTM1 and CYP1A1 metabolic genes: A case-control study in oncohematological diseases

AIM To analyze the association between oncohematological diseases and GSTT1 /GSTM1 /CYP1A1 polymorphisms, dietary habits and smoking, in an argentine hospitalbased case-control study.METHODS This hospital-based case-control study involved 125 patients with oncohematological diseases and 310 control subjects. A questionnaire was used to obtain sociodemographic data and information about habits. Blood samples were collected, and DNA was extracted using salting out methods. Deletions in GSTT1 and GSTM1 (null genotypes) were addressed by PCR. CYP1A1 MspI polymorphism was detected by PCR-RFLP. Odds ratio(OR) and 95%CI were calculated to estimate the association between each variable studied and oncohematological disease.RESULTS Women showed lower risk of disease compared to men(OR 0.52, 95%CI: 0.34-0.82, P = 0.003). Higher levels of education(> 12 years) were significantly associated with an increased risk, compared to complete primary school or less(OR 3.68, 95%CI: 1.82-7.40, P < 0.001 adjusted for age and sex). With respect to tobacco, none of the smoking categories showed association with oncohematological diseases. Regarding dietary habits, consumption of grilled/barbecued meat 3 or more times per month showed significant association with an increased risk of disease(OR 1.72, 95%CI: 1.08-2.75, P = 0.02). Daily consumption of coffee also was associated with an increased risk(OR 1.77, 95%CI: 1.03-3.03, P = 0.03). Results for GSTT1, GSTM1 and CYP1A1 polymorphisms showed no significant association with oncohematological diseases. When analyzing the interaction between polymorphisms and tobacco smoking or dietary habits, no statistically significant associations that modify disease risk were found. CONCLUSION We reported an increased risk of oncohematological diseases associated with meat and coffee intake. We did not find significant associations between genetic polymorphisms and blood cancer.

The Effect of the Cyp19a1 Gene Methylation Modification on Temperature-dependent Sex Determination of Reeves' Turtle(Mauremys reevesii)

Temperature-dependent sex determination（TSD） is a type of environmental sex determination in which the sex of the embryos depends on the ambient temperature; however,the molecular mechanisms governing this process remain unknown.Aromatase,encoded by the cyp19a1 gene,which converts androgens into estrogens in animals,was considered to be the key gene for TSD.In this study,the 5＇-flanking region of the cyp19a1 gene in Reeves＇ turtle（Mauremys reevesii） was cloned,and the promoter region was identified using the luciferase reporter assay.Then the eggs of Reeves＇ turtle were incubated at different temperatures（26°C： male-biased temperature; 29°C： non-sex-biased temperature and 32°C： female-biased temperature）.During the thermosensitive period,the adrenal kidney gonad complexes（AKG） were sampled.DNA methylation analysis of the AKG samples showed that the promoter region of the cyp19a1 gene was significantly de-methylated in the female-biased temperature regime（P＆lt;0.01）.Quantitative analysis of the cyp19a1 gene and estrogen by q PCR and Elisa assay showed that the expression level of the cyp19a1 gene and estrogen content were both upregulated significantly at the female-biased temperature（P＆lt;0.01）.These results indicated that the de-methylation response of the cyp19a1 gene to incubation temperature,especially at the female-biased temperature,could lead to temperature-specific expression of aromatase and increased estrogen levels,which may further determine gonadal development in Reeves＇ turtle.These findings provide insights into the genetic mechanisms underlying TSD.

Molecular cloning and expression patterns of the cholesterol side chain cleavage enzyme(CYP11A1) gene during the reproductive cycle in goose(Anas cygnoides)

Background： CYP11A1, a gene belonging to the family 11 of cytochrome P450, encodes a crucial steroidogenic enzyme that catalyzes the initial step in the production of all classes of steroids. Many studies show that CYP11A1 plays a role in ovary function. However, the role of CYP11A1 in goose reproductive cycle remains largely unknown.Results： In this study, full-length CYP11A1 c DNA of Zhedong goose was obtained using reverse transcription polymerase chain reaction（RT-PCR） and rapid amplification of c DNA ends（RACE）. The c DNA consisted of a 96-base pair（bp） 5′untranslated region（UTR）, a 179-bp 3′UTR and a 1509-bp open reading frame. The open reading frame encodes a putative 503 amino acid protein that shares high homology with CYP11A1 of other birds. The amino acid sequence possesses conserved domains of the P450 superfamily, which include the steroid-binding domain and the heme-binding region. Real-time quantitative polymerase chain reaction（q PCR） analysis revealed CYP11A1 mR NA was expressed ubiquitously in every Zhedong goose tissue analyzed, including the heart, liver, glandular stomach,lung, spleen, kidney, intestinum tenue, intestinum crassum, cerebrum, cerebellum, muscle, oviduct, pituitary,hypothalamus and ovary.. The relatively low levels of CYP11A1 m RNA were detected in pituitary, ovary and oviduct tissues at ovulation when compared with levels at oviposition. Interestingly, higher expression was observed in ovary and oviduct tissues during brooding. Lastly, higher m RNA expression of Yangzhou geese was detected during the ovulation period than that of Zhedong geese.Conclusions： Our findings reveal the sequence characterization and expression patterns of the CYP11A1 gene during the goose reproductive cycle, which may provides correlative evidence that CYP11A1 expression is important in reproduction activity.

Associations of Polymorphisms of the <i>CYP</i>1<i>A</i>1 and <i>CYP</i>1<i>B</i>1 Cytochrome P450 Genes with Breast Cancer in Kazakhstan

Associations of polymorphisms in rs4646903 site of CYP1A1 and rs1056836 site of CYP1B1 genes with the breast cancer (BC) were studied in two main ethnic groups of Kazakhstan Republic (Kazakhs and Russians). Total number of BC patients was 181, controls—397. The statistically significant differences were revealed in allele frequencies (χ2 = 5.93, р = 0.004) and in genotypes distribution (χ2 = 8.71, р = 0.015) in rs4646903 site of CYP1A1 gene in Kazakh but not in Russian group. The study of CYP1В1 rs1056836 site demonstrated differences in genotype distributions (χ2 = 7.48, р = 0.023) between BC patients and controls in Russian but not in Kazakh ethnic group.

亲环素基因GhCYP1在陆地棉中的过量表达及耐盐性分析

通过农杆菌介导法将亲环素基因GhCYP1导入陆地棉棉花栽培品种中棉35中,经过卡那霉素抗性选择、PCR检测和系统选育获得5个不同转基因纯合系.在温室盆栽条件下,于3片真叶期对转基因棉花纯合系和非转基因对照进行200 mmol/L NaCl胁迫处理20d.结果表明,转GhCYP1基因棉花株系比对照长势强,株高比对照提高2～5cm,地上部分单株鲜质量比对照增加7.1％～12.4％,抗氧化物酶SOD,POD,CAT等的活性以及叶绿素含量显著高于对照.说明过量表达GhCYP1基因提高了陆地棉对盐碱的抗性.

代谢酶基因CYP 1A1与广西胃癌遗传易感性的关系

目的探讨代谢酶基因CYP 1A1MSP1多态性与广西地区汉族、壮族人群胃癌遗传易感性之间相关性。方法采用PCR-RFLP技术检测广西地区汉族、壮族人群中121例胃癌患者和138例健康人CYP1A1MSP1多态性的分布频率,分析其与广西地区汉族、壮族人群胃癌遗传易感性的相关性及与吸烟、饮酒在胃癌易感性中的相互作用。结果 (1)CYP 1A1MSP1三种基因型(m1/m1、m1/m2、m2/m2)分布频率在两组间比较差异无统计学意义(χ2=0.901,P=0.342);(2)携带CYP1A1MSP1突变m2基因型的个体较携带m1/m1基因型的个体患胃癌的风险增加(OR=1.509),但差异无统计学意义(P=0.342)。结论单独的CYP 1A1基因MSP1多态性与广西地区汉族、壮族人群胃癌易感性无明显相关性。

PPAR-γC161T和CYPⅡE1基因多态性与脂肪肝中医证型的关系

目的探讨PPARγC161-T、CYPⅡE1基因多态性与脂肪肝中医证型的关系。方法将183例脂肪肝患者辨证分为瘀证组、痰证组和无证可辨组,检测183例患者及40例健康者的身体质量指数(BMI)、腰围、血压、肝肾功能、血脂、空腹血糖(FBG)、HOMA–IR(稳态胰岛素抵抗指数),检测其PPARγC161-T、CYPⅡE1的基因多态性及等位基因频率。结果瘀证组和痰证组总甘油三脂(TG)(P<0.01)、总胆固醇(TC)(P<0.05)、低密度脂蛋白(LDL-C)(P<0.05)均高于无证可辨组。PPAR-γC161-T基因型频率及等位基因频率在脂肪肝各证型组与对照组中无统计学差异(P>0.05)。CYPⅡE1基因型频率及等位基因频率在脂肪肝各证型组与对照组中有统计学差异(P<0.05)。结论脂质代谢紊乱及相关改变与脂肪肝痰、瘀证密切相关。CYPⅡE1基因多态性可能在脂肪肝痰瘀证的形成以及转化中起着决定性的作用。

蒙古族与汉族人群GSTM3及CYP1A1基因多态性的研究

目的研究内蒙古地区蒙、汉族人群GSTM3和CYP1A1 Exon7基因多态性。方法采用聚合酶链式反应-限制性片断长度多态性(PCR-RFLP)和等位基因特异性扩增(ASA)技术分析内蒙古地区412例健康蒙古族人和436例健康汉族人的GSTM3及CYP1A1 Exon7基因多态性及基因型分布频率。结果内蒙古地区蒙、汉族人群GSTM3基因型之间的分布频率具有显著性差异(P<0.05),CYP1A1 Exon7基因型分布频率无显著性差异(P>0.05)。结论 GSTM3基因型频率在内蒙古地区健康蒙、汉族人群中的分布具有显著差异,而CYP1A1 Exon7基因型分布频率无显著性差异。

PCR和探针自我杂交──Northern Blot方法对二甲基苯蒽诱导的P_（450）Cyp^（1b－1）mRNA的定量

通过ＰＣＲ和探针自我杂交──ＮｏｒｔｈｅｒｎＢｌｏｔ方法对０．３μＭ二甲基苯蒽（ＤＭＢＡ）诱导的２×１０６个小鼠胚胎成纤维细胞１０Ｔ１／２产生的Ｐ４５０Ｃｙｐ１ｂ－１ｍＲＮＡ进行了定量。在这个方法中，与Ｐ４５０Ｃｙｐ１ｂ－１ｍＲＮＡ相同的特定的单链ＣＤＮＡ探针片断通过ＰＣＲ合成。而双链探针中的一条链与单链探针和Ｐ４５０Ｃｙｐ１ｂ－１ｍＲＮＡ的特定片断是相同的。双链探针上的另一条链（Ｈ链）则与单链探针和Ｐ４５０Ｃｙｐ１ｂ－１ｍＲＮＡ的特定片断是互补的。用３２Ｐ－ｄＣＴＰ标记Ｈ链后，它就能够与单链探针和Ｐ４５０Ｃｙｐ１ｂ－１ｍＲＮＡ同时进行杂交。这样，就能用已知浓度的单链探针作为标准物制做标准工作曲线和对Ｐ４５０Ｃｙｐ１ｂ－１ｍＲＮＡ的含量进行定量。经测定，在０．３μＭＤＭＢＡ分别于１、２、３、４、６小时对２×１０６个细胞进行诱导后，２×１０６个细胞在不同时间产生的Ｐ４５０Ｃｙｐ１ｂ－１ｍＲＮＡ分别为０．００９８、０．０１３５、０．０１７４、０．０１９０和０．０１２０μＭ。

CYP2E1 RsaⅠpolymorphism impacts on risk of colorectal cancer association with smoking and alcohol drinking

AIM： To investigate associations between the Rsa I polymorphism of CYP2E1 and risk of colorectal cancer. METHODS： A case-control study was conducted with 315 colorectal cancer cases （105 colon, 210 rectal） and 439 population-based controls in Jiangsu Province of China. Genomic DNA samples were assayed for restriction fragment length polymorphisms in CYP2E1 by PCR amplification followed by digestion with Rsa I. Information on smoking and alcohol drinking was collected using a questionnaire. Odds ratios （ORs） were estimated with an unconditional logistic model. RESULTS： The proportional distribution of the CYP2E1 Rsa I c1/c1, c1/c2 and c2/c2 genotypes were 61.4%, 35.6% and 3.0% in controls, 60.6%, 33.7% and 5.8% in colon cancer cases, and 58.4%, 34.0% and 7.7% in rectal cancer cases, respectively. A significant differencewas noted between controls and rectal cancer cases （P = 0.029）, the c2/c2 genotype being associated with elevated OR （adjusted age, sex and status of the smoking and alcohol drinking） for rectal cancer （1.64, 95% CI, 1.12-2.41, vs cl allele carriers）, but not for colon cancer. In interaction analysis between the CYP2E1 Rsa I genotype and smoking and drinking habits, we found a significant cooperative action between the c2/c2 genotype and alcohol drinking in the sex-, age-adjusted ORs for both colon （4.74, 95% CI, 1.10-20.40） and rectal （5.75, 95% CI, 1.65-20.05） cancers. Among nonsmokers, the CYP2E1 Rsa I c2/c2 genotype was also associated with elevated ORs in the two sites （1.95, 95% CI, 0.99-3.86 and 2.30, 95% CI, 1.32-3.99）. CONCLUSION： The results of the present study suggest that the CYP2E1 c2/c2 genotype increases susceptibility to rectal cancer and the gene-environmental interactions between the CYP2E1 polymorphism and smoking or alcohol drinking exist for colorectal neoplasia in general.

Relationship between CYP1A1 polymorphisms and invasion and metastasis of breast cancer

Objective:To investigate the relationship between CYPIA1 genetic polymorphisms and the invasion and metastasis of breast cancer.Methods:The CYP1A1 gene polymorphism(an T-C transversion at nucleotide position 3801)was detected by the polymerase chain reaction and restriction fragment length polymorphism in 80 cases with breast cancer and 60 samples of normal breast tissue.The difference in genotypic distribution frequency between the groups,the correlation between the genotypes and the factors related to prognosis were analyzed.Results:The incidence of homozygous and variant genotypes had no difference between the breast cancer group and controls group(P=0.746).The proportion of variant genotype increased as clinical stage(P=0.006)advanced,as well as with increased numbers of lymph node metastases(P=0.010).Conclusions:In patients with breast cancer there is a correlation between the CYP1A1 CC allele and some factors indicating poor prognosis,including more lymph node metastases as well as a more advanced clinical stage.

Cytochrome P450 2E1 genetic polymorphism and gastric cancer in Changle,Fujian Province

AIM: Genetic polymorphism in enzymes of carcinogen metabolism has been found to have the influence on the susceptibility to cancer. Cytochrome P450 2E1 (CYP2E1) is considered to play an important role in the metabolic activation of procarcinogens such as N-nitrosoamines and low molecular weight organic compounds. The purpose of this study is to determine whether CYP450 2E1 polymorphisms are associated with risks of gastric cancer. METHODS: We conducted a population based case-control study in Changle county, Fujian Province, a high-risk region of gastric cancer in China. Ninety-one incident gastric cancer patients and ninety-four healthy controls were included in our study. Datas including demographic characteristics, diet intake, and alcohol and tobacco consumption of individuals in our study were completed by a standardized questionnaire.PCR-RFLP revealed three genotypes:heterozygote (C1/C2) and two homozygotes (C1/C1 and C2/C2) in CYP2E1. RESULTS: The frequency of variant genotypes (C1/C2 and C2/C2) in gastric cancer cases and controls was 36.3% and 24.5%, respectively. The rare homozygous C2/C2 genotype was found in 6 individuals in gastric cancer group(6.6%), whereas there was only one in the control group (1.1%). However, there was no statistically significant difference between the two groups (two-tailed Fisher's exact test P=0.066). Individuals in gastric cancer group were more likely to carry genotype C1/C2 (odds ratio, OR=1.50) and C2/C2 (OR=7.34) than individuals in control group (chi(2) =4.597, for trend P=0.032). The frequencies of genotypes with the C2 allele (C1/C2 and C2/C2 genotypes) were compared with those of genotypes without C2 allele (C1/C1 genotype) among individuals in gastric cancer group and control group according to the pattern of gastric cancer risk factors. The results show that individuals who exposed to these gastric cancer risk factors and carry the C2 allele seemed to have a higher risk of developing gastric cancer. CONCLUSION: Polymorphism of CYP2E1 gene may have some effect in the development of gastric cancer in Changle county, Fujian Province.

Metabolism of Terephthalic Acid and Its Effects on CYP4B1 Induction

Objective To investgate the metabolism of terephthalic acid （TPA） in rats and its mechanism. Methods Metabolism was evaluated by incubating sodium terephthalate （NaTPA） with rat normal liver microsomes, or with microsomes pretreated by phenobarbital sodium, or with 3-methycholanthrene, or with diet control following a NADPH-generating system. The determination was performed by high performance liquid chromatography （HPLC）, and the mutagenic activation was analyzed by umu tester strain Salmonella typhimurium NM2009. Expression of CYP4B 1 mRNA was detected by RT-PCR. Results The amount of NaTPA （12.5-200 μmol-L^1） detected by HPLC did not decrease in microsomes induced by NADPH-generating system. Incubation of TPA （0.025-0.1 mmol-L^-1） with induced or noninduced liver microsomes in an NM2009 umu response system did not show any mutagenic activation. TPA exposure increased the expression of CYP4B1 mRNA in rat liver, kidney, and bladder. Contusion Lack of metabolism of TPA in liver and negative genotoxic data from NM2009 study are consistent with other previous short-term tests, suggesting that the carcinogenesis in TPA feeding animals is not directly interfered with TPA itself and/or its metabolites.

Increased Risk of Acute Myeloid Leukemia in Patients with CYP1A1 Polymorphisms

Acute Myeloid Leukemia (AML) is a group of genetically diverse hematopoietic malignancies arising from cell progenitors developing in the myeloid pathway or from primitive stem cells. Genetic susceptibility of AML may account for an increased risk of AML due to partial metabolism of or biocativation of carcinogens. Chemical compounds are metabolized by a two-tiered phase detoxifying system. Polymorphisms in these pathways may lead to DNA damage and development of AML. We determined the frequencies of carcinogen metabolism gene polymorphisms (CYP1A1, del{GSTM1} and del{GSTT1}) in a case control-study based on polymorphism analysis. Fifty-eight consecutively AML patients (median age 62 years) and 174 sex and age-matched control group were assessed by a PCR-RFLP assay. There were 51 de novo and 7 secondary AML. CYP1A1*2A and CYP1A1*2C polymorphisms were more frequent in CG than AML p 0.001 and in contrast, CYP1A1*3 and CYP1A1*4 were more frequent in AML than CG p 0.001. There were no differences in del{GSTM1} neither del{GSTT1} between AML and CG (p = 0.999 and p = 0.539). Odds ratio for AML in patients harboring CYP1A1*3 was 2.36 (95% CI 1.2 - 4.5), 2.38 for CYP1A1*4 (95% CI 0.8 - 6.8). Adjusted OR was 2.63 for CYP1A1*3 (95% CI 1.4 - 5.1) and 2.66 for CYP1A1*4 (95% CI 0.9 - 7.8). In the multivariate analysis CYP1A1*3 polymorphism was a risk factor for AML with an OR for 3.99 (95%CI 1.9 - 8.6). To the best of our knowledge this is the first study to show that CYP1A1*3 heterozygous genotypes increase the risk of AML. Our data support that inherited absence of this carcinogen detoxification pathway may be an important determinant of AML.

Progressive ataxia of cerebrotendinous xanthomatosis with a rare c.255+1G>T splice site mutation:A case report

BACKGROUND Cerebrotendinous xanthomatosis is an autosomal recessive disorder of lipid metabolism caused by the mutation of the CYP27A1 gene encoding sterol 27-hydroxylase,an essential enzyme for the conversion of cholesterol to chenodeoxycholic and cholic acids.Cerebrotendinous xanthomatosis is a rare neurological dis-ease with a wide range of clinical symptoms that are easily misdiagnosed.CASE SUMMARY Here we report the clinical,biochemical,and molecular characterization of a 33-year-old female patient with cerebrotendinous xanthomatosis.The patient developed ataxia and had the typical symptoms of juvenile cataracts,tendon xanthomata,and progressive nervous system dysfunction.Magnetic resonance imaging of the brain revealed bilateral dentate nucleus lesions and white matter abnormalities.This patient was misdiagnosed for 2 years resulting in severe neurological complications.After 2 years of chenodeoxycholic acid treatment,she still presented with ataxia and dysarthria.The pathogenic sites of CYP27A1 were identified as c.255+1G>T and c.1263+1G>T,which were both caused by shear denaturation.CONCLUSION Cerebrotendinous xanthomatosis requires a multidisciplinary diagnosis that must be made early to avoid progressive neurological degeneration.c.1263+1G>T is a known mutation,but c.255+1G>T is a rare mutation site.

The microRNA miR-184 regulates the CYP303A1 transcript level to control molting of Locusta migratoria

Cytochrome P450 monooxygenases(CYPs)play essential physiological functions in insects.CYP303A1 is highly conserved in insect species studied to date,and shows an indispensable role for adult eclosion in both Locusta migratoria and Drosophila melanogaster.However,how CYP303A1 is regulated to control insect developmental processes remains uninvestigated.In this study,we discovered functional binding sites for miR-184 in the coding sequence of LmCYP303A1.The luciferase reporter assay showed that miR-184 could target LmCYP303A1 and regulate its expression in vitro.Furthermore,overexpression of miR-184 through microinjection of agomir to locusts reduced the transcripts of LmCYP303A1 and led to abnormal molting,which is similar to the phenotype of silencing LmCYP303A1 by direct injection of dsLmCYP303A1 to locusts.Meanwhile,down-regulation of miR-184 by injection of antagomir increased the LmCYP303A1 transcript and caused molting defects.These findings suggested that miR-184 could target LmCYP303A1 to regulate the molting process in L.migratoria,which might be considered as a novel target for pest control.

Preferential expression of cytochrome CYP CYP2R1 but not CYP1B1 in human cord blood hematopoietic stem and progenitor cells

Cytochrome P450(CYP)enzymes metabolize numerous endogenous substrates,such as retinoids,androgens,estrogens and vitamin D,that can modulate important cellular processes,including proliferation,differentiation and apoptosis.The aim of this study is to characterize the expression of CYP genes in CD34+human cord blood hematopoietic stem and early progenitor cells(CBHSPCs)as a first step toward assessment of the potential biological functions of CYP enzymes in regulating the expansion or differentiation of these cells.CD34+CBHSPCs were purified from umbilical cord blood via antibody affinity chromatography.Purity of CD34+CBHSPCs was assessed using fluorescence-activated cell sorting.RNA was isolated from purified CD34+CBHSPCs and total mononuclear cells(MNCs)for RNA-PCR analysis of CYP expression.Fourteen human CYPs were detected in the initial screening with qualitative RT-PCR in CD34+CBHSPCs.Further quantitative RNA-PCR analysis of the detected CYP transcripts yielded evidence for preferential expression of CYP2R1 in CD34+CBHSPCs relative to MNCs;and for greater expression of CYP1B1 in MNCs relative to CD34+CBHSPCs.These findings provide the basis for further studies on possible functions of CYP2R1 and CYP1B1 in CBHSPCs'proliferation and/or differentiation and their potential utility as targets for drugs designed to modulate CD34+CBHSPC expansion or differentiation.