Novel single nucleotide polymorphism of UGT1A7 gene in Japanese

We sequenced exon 1 of the UDP-glucuronosyltransferase (UGT) 1A7 gene from 52 Japanese cancer patients. Four single nucleotide polymorphisms (SNPs) were found. Three of them caused UGT1A7*2 and UGT1A7*3. A novel SNP (98973G>C) causing amino acid substitution (Ser141Cys) was found. The sequence is as follows: SNP, 050824FujitaK002; Gene Name, UGT1A7; Accession Number, AF297093; Length, 25 bases; 5'-TAAAGGAGAGTTG/CTTTTGATGCAGT-3'. One out of 52 cancer patients was heterozygous for the variant allele, resulting in the allele frequency of 0.96%. The patient did not possess UGT1A7*2 or UGT1A7*3.     

Three novel single nucleotide polymorphisms in UGT1A9

Three novel single nucleotide polymorphisms (SNPs) were found in the UDP-glucuronosyltransferase (UGT) 1A9 gene from 97 Japanese subjects (47 cancer patients and 50 cardiovascular disease patients). The detected SNPs were as follows: 1) SNP, MPJ6_U1A006; GENE NAME, UGT1A9; ACCESSION NUMBER, AF297093; LENGTH, 25 bases; 5'-AATTCTCTTAGGG/TTTCTCAGATGCC-3'. 2) SNP, MPJ6_U1A007; GENE NAME, UGT1A9; ACCESSION NUMBER, AF297093; LENGTH, 25 bases; 5'-TGTTACGGAGTAT/GGATCTCTACAGC-3'. 3) SNP, MPJ6_U1A031; GENE NAME, UGT1A9; ACCESSION NUMBER, AF297093; LENGTH, 25 bases; 5'-ACTCATTCTCAGG/AGGGCATGAGGTG-3'. All three SNPs were located in exon 1 with frequencies of 0.036 for MPJ6_U1A006, and 0.005 for MPJ6_U1A007 and MPJ6_U1A031. SNP MPJ6_U1A007 (726T>G) results in formation of a termination codon TAG (Y242X). The other two SNPs, MPJ6_U1A006 (588G>T) and MPJ6_U1A031 (153G>A), result in synonymous changes (G196G and R51R, respectively).     

Novel single nucleotide polymorphism of the CYP2A13 gene in Japanese individuals

Cytochrome P450 2A13 (CYP2A13) is a human CYP enzyme that is selectively expressed in the respiratory tract. It plays an active role in the metabolic activation of a tobacco-specific procarcinogen. In this study, the entire coding sequence and the exon-intron junctions of the CYP2A13 gene obtained from 395 Japanese individuals were screened for genetic polymorphisms. Eight genetic polymorphisms were found, of which seven gave rise to known variant alleles: CYP2A13*2, CYP2A13*3, CYP2A13*4, CYP2A13*6, and CYP2A13*7. We identified a novel single nucleotide polymorphism (SNP), 5792T>C, in exon 7 that caused an amino acid substitution (Ile331Thr). One of the 395 individuals included in the study was heterozygous for the variant allele, and therefore, the frequency of the allele in the study population was 0.13%.     

A novel single nucleotide polymorphism of the human methylenetetrahydrofolate reductase gene in Japanese individuals

The genetic polymorphisms of methylenetetrahydrofolate reductase (MTHFR) have been associated with increased toxicity of methotrexate (MTX), a folic acid antagonist that is widely used to treat cancer and immunosuppressive disorders such as rheumatoid arthritis. In this study, we analyzed all the exons and exon/intron junctions of the MTHFR gene from 200 Japanese individuals. We detected a novel single nucleotide polymorphism (SNP) 148C>T (Arg46Trp) in exon 1. The allele frequency of this polymorphism in the Japanese population appears to be extremely low (0.25%).     

Three novel single nucleotide polymorphisms in UGT1A10

Three novel single nucleotide polymorphisms (SNPs) were found in the UDP-glucuronosyltransferase (UGT) 1A10 gene from 24 Japanese patients with various cancers who were administered the anti-tumor drug, irinotecan (CPT-11). The detected SNPs were as follows: 1) SNP, MPJ6_U1A003; GENE NAME, UGT1A10; ACCESSION NUMBER, AF297093; LENGTH, 25 bases; 5'-CAGATGCCATGAC/TTTTCAAGGAGAG-3'. 2) SNP, MPJ6_U1A004; GENE NAME, UGT1A10; ACCESSION NUMBER, AF297093; LENGTH, 25 bases; 5'-CCTAGAAATAGCC/TTCTGAAATTCTC-3'. 3) SNP, MPJ6_U1A030; GENE NAME, UGT1A10; ACCESSION NUMBER, AF297093; LENGTH, 25 bases; 5'-GGTTGTAGTCATG/ACCAGAGGTGAGT-3' All the three SNPs were located in exon 1 and their frequencies were all 0.021. Among these SNPs, MPJ6_U1A003 and U1A030 resulted in amino acid alterations, T202I and M59I, respectively. The third SNP, MPJ6_U1A004, introduced a synonymous amino acid change (A231A).     

A novel polymorphism in the promoter region of human UGT1A9 gene (UGT1A9*22) and its effects on the transcriptional activity

The human UDP-glucuronosyltransferase, UGT1A9, catalyses glucuronidations of various endobiotics and xenobiotics. In the present study, all exons, exon-intron junctions, and the 5'-flanking region (-273 bp) of the UGT1A9 gene in a Japanese subject were sequenced. One base insertion of thymidine in a promoter region of the UGT1A9 gene resulting in A(T)10AT was identified compared to the reference sequence of AF297093 (A(T)9AT). The allele was termed UGT1A9*22. A polymerase chain reaction-single strand conformation polymorphism method was developed to genotype the allele. The allele frequencies of the mutation in 87 Japanese, 50 Caucasian and 50 African-American subjects were 60%, 39% and 44%, respectively. The significance of the polymorphism was investigated by the construction of luciferase reporter plasmids containing 170 bp of the 5'-flanking region of the gene transfected into human hepatoma HepG2 cells. The luciferase activity of the promoter construct containing the A(T)10AT sequence was 2.6-fold higher than that of the construct containing the A(T)9AT sequence. In conclusion, the mutant allele with one base insertion in the promoter region of the UGT1A9 gene would alter the level of enzyme expression and the metabolism of those drugs that are substrates of UGT1A9.     

贵州省水族人群XRCC1基因单核苷酸多态性及其组合型研究

目的 分析水族人群DNA修复基因XRCC1 C26304T、G27466A与G28152A单核苷酸多态性及其等位基因频率与组合分布特征.方法 获取自然人群个体水族197例血白细胞基因组DNA,利用PCR扩增限制性酶切法(PCR-RFLP)检测XRCC1 C26304T、G27466A与G28152ASNPs.结果 XRCC1 C26304T SNPs基因型分布CC、CT、TT分别为57.4%、33.5%、9.1%,C、T等位基因频率分别为74.1%、25.9%.XRCC1 G27466A SNPs基因型分布GG、GA、AA分别为79.7%、18.9%、1.5%,G、A等位基因频率分别为89.1%、10.9%.XRCC1 G28152 A SNPs基因型分布GG、GA、AA分别为49.2%、43.7%、7.1%,G、A等住基因频率分别为71.1%、28.9%.XRCC1基因在C26304T和G28152A两位点的SNPs组合基因型频率较高的为CC GA 53例(26.9%);C26304T和G274 66A两位点的SNPs组合基因型频率较高的为CC GG 84例(42.6%);G28152A和G27466 A两位点的SNPs组合基因型频率较高的为GA GG 73例(37.1%).结论 本研究揭示了水族人群XRCC1基因3位点的单核苷酸多态性、基因型及其组合分布特征,为进一步研究该基因SNPs与其生理功能和疾病的关系提供基础资料.     

Glucuronidation of flavonoids by recombinant UGT1A3 and UGT1A9

Flavonoids are highlighted for their potential roles in the prevention of oxidative stress-associated diseases. Their metabolisms in vivo, such as glucuronidation, are the key points to determine their health beneficial properties. In this paper, we tested the glucuronidation of nineteen flavonoids by both recombinant human UGT1A3 and UGT1A9. Eleven compounds could be catalyzed by both enzymes. In general, both enzymes showed moderate to high catalyzing activity to most flavonoid aglycones, while the catalyzing efficiency changed with structures. Each flavonoid produced more than one monoglucuronide with no diglucuronide detected by liquid chromatography-mass spectrometry (LC-MS). Enzymatic kinetic analysis indicated that the catalyzing efficiency (Vmax/Km) of UGT1A9 was higher than that of UGT1A3, suggesting its important role in flavonoid glucuronidation. Both human UGT1A3 and UGT1A9 preferred flavonoid aglycone to flavonoid glycoside, and their metabolism to arabinoside was stronger than to other glycosides. Of the flavonoids studied, it is the first time to report isorhamnetin, morin, silybin, kaempferol, daidzein, quercetin-3',4'-OCHO-, quercetin xylopyranoside and avicularin as substrates of UGT1A3. Apigenin, morin, daidzein, quercetin-3',4'-OCHO-, quercetin xylopyranoside and avicularin were the newly reported substrates of UGT1A9.     

ORAI1基因rs3741596单核苷酸多态性与川崎病的关系

摘要： 目的 探讨钙释放激活钙调节蛋白1 （CRACM1/ORAI1） 基因rs3741596位点单核苷酸多态性 （SNP） 与川崎病 （KD） 易感性及KD并发冠状动脉病变 （CALs） 是否相关。方法 将确诊的46例KD患儿 （病例组） 和25例健康儿童（对照组） 纳入本研究, 并根据是否出现CALs将病例组分为CAL组 （20例） 和无CAL （NCAL） 组 （26例）。应用聚合酶链反应 （PCR） 技术联合基因直接测序技术检测所有研究对象ORAI1基因rs3741596位点SNP, 并进行统计学分析。结果 所有研究对象均检测到ORAI1基因rs3741596位点SNP, 其基因型分布及等位基因频率在病例组及对照组间差异无统计学意义 （χ2 分别为0.712和0.499, 均P＞0.05）, 而在CAL组和NCAL组间差异有统计学意义 （χ2 分别为 6.524 和 6.891, 均 P＜0.05）; 携带 G 等位基因使 KD 患儿并发生 CALs 的危险性增加（OR=5.444, 95%CI： 1.386~ 21.380）。结论 ORAI1基因rs3741596位点SNP可能与KD易感性无相关性, 但与KD并发CALs易感性相关。     

Influence of UGT1A7 and UGT1A9 intronic I399 genetic polymorphisms on mycophenolic acid pharmacokinetics in Japanese renal transplant recipients

UGT1A7 and UGT1A9 are uridine diphosphate-glucuronosyltransferase isoforms involved in the glucuronidation of mycophenolic acid (MPA). The aim of this study was to elucidate MPA pharmacokinetics in UGT1A7 and UGT1A9 intronic I399 genotypes in Japanese adult renal transplant recipients. Eighty recipients were given repeated doses of combination immunosuppressive therapy consisting of mycophenolate mofetil and tacrolimus every 12 hours at a designated time (9:00 am and 9:00 pm). On day 28 after renal transplantation, plasma MPA concentrations were measured by high-performance liquid chromatography. All patients had UGT1A9 98TT/-275TT/-2152CC and UGT1A10 177GG/605CC genotypes. The UGT1A7*1/*1, *1/*2, *1/*3, *2/*3, and *3/*3 genotypes were detected in 35 (43.8%), five (6.2%), 28 (35.0%), eight (10.0%), and four (5.0%) patients, respectively, and the UGT1A9 I399C/C, C/T, and T/T genotypes were detected in 12 (15.0%), 33 (41.2%), and 35 (43.8%) patients of the 80 Japanese recipients. There were no significant differences in MPA pharmacokinetics among UGT1A7 or UGT1A9 intronic I399 genotype groups. The mean dose-adjusted area under the plasma concentration-time curve from zero to 12 hours (AUC0-12) of MPA in UGT1A7*1/*1, *1/*2, *1/*3, *2/*3, and *3/*3 were 95, 98, 99, 88, and 86 ng.h/mL/mg, respectively (P = 0.9475). The mean dose-adjusted AUC0-12 of MPA in UGT1A9 I399C/C, C/T, and T/T were 87, 99, and 95 ng.h/mL/mg, respectively (P = 0.6937). The dose-adjusted trough levels of MPA in UGT1A9 I399C/C, C/T, and T/T were 5.4, 5.5, and 4.7 ng/mL/mg (P = 0.5845). Although UGT1A7*3 and UGT1A9 I399C/C are known to have low-activity variants when studied in vitro, they do not have reduced in vivo MPA glucuronidation activity. UGT1A7 and UGT1A9 I399 polymorphisms do not contribute to interindividual differences in MPA pharmacokinetics.     

焦磷酸测序技术检测UGT1A3和UGT2B7在中国汉族人群中的基因多态性

目的建立焦磷酸测序技术(pyrosequencing)研究二相代谢酶UGT1A3和UGT2B7基因多态性在中国汉族人群中的分布。方法应用带生物素标记扩增引物并经PCR扩增和Beads分离,制备UGT1A3和UGT2B7焦磷酸测序单倍摸板。在PYroMarkID焦磷酸测序上进行焦磷酸测序,检测233血样的DNA标本的17个SNP位点,以确定血样DNA标本的的基因型。结果 233例血样的DNA标本中,UGT1A3等位基因有9种表型,分别为UGT1A3*1*1、UGT1A3*1*2、UGT1A3*1*3、UGT1A3*1*4、UGT1A3*1*5、UGT1A3*2*3、UGT1A3*2*4、UGT1A3*3*3和UGT1A3*3*5。UGT2B7-1和UGT2B7-2各有3种基因型,分别为G/G型、G/T型、T/T和C/C型、C/T型、T/T型。结论我国汉族人群中UGT1A3和UGT2B7基因突变较高。     

UGT1A1基因多态性对宝安地区女性妊娠期甲状腺功能减退的临床价值

目的:探究尿苷二磷酸葡萄糖醛酸转移酶1A1（UGT1A1）基因多态性对宝安地区女性妊娠期甲状腺功能减退的临床价值。方法:以2021年1月至2022年2月就诊于深圳市宝安区石岩人民医院妊娠期甲状腺功能减退的女性100例为观察组,同时选取同期产检甲状腺功能正常的健康女性100例为对照组。取所有研究对象的空腹静脉血,进行RN...     

Novel nonsynonymous polymorphisms of the CYP1A1 gene in Japanese

We found five novel nonsynonymous polymorphisms of the human CYP1A1 gene from Japanese individuals. The five single nucleotide polymorphisms (SNP) in exon 7 (2346_2347 ins T, 2414T>A, 2461C>T, 2500C>T and 2546C>G causing premature stop codon, Ile(448)Asn, Arg(464)Cys, and Arg(477)Trp and Pro(492)Arg, respectively) were as follows:SNP, 030212Saito001; GENE NAME, CYP1A1; ACCESSION NUMBER, X02612; LENGTH, 25 base; 5'-GTCAACCCATCT-/TGAGTTCCTACCT-3'.SNP, 030212Saito002; GENE NAME, CYP1A1; ACCESSION NUMBER, X02612; LENGTH, 25 base; 5'-GTGAGAAGGTGAT/ATATCTTTGGCAT-3'.SNP, 030212Saito003; GENE NAME, CYP1A1; ACCESSION NUMBER, X02612; LENGTH, 25 base; 5'-GAGACCGTTGCCC/TGCTGGGAGGTCT-3'.SNP, 030212Saito004; GENE NAME, CYP1A1; ACCESSION NUMBER, X02612; LENGTH, 25 base; 5'-ATCCTGCTGCAAC/TGGGTGGAATTCA-3'.SNP, 030212Saito005; GENE NAME, CYP1A1; ACCESSION NUMBER, X02612; LENGTH, 25 base; 5'-TGGACATGACCCC/GCATCTATGGGCT-3'.     

Structure-metabolism relationships for the glucuronidation of flavonoids by UGT1A3 and UGT1A9

Objectives This study tries to find structure–metabolism relationships between flavonoids and human UGT1A3 and UGT1A9. Methods The glucuronidation of flavonoids was studied with recombinant UGT1A3 and UGT1A9, and the glucuronidation activity was determined by HPLC. Key findings Of the flavonoids studied, it was shown for the first time that baicalein, quercetin‐3‐OCH₂OCH₃, quercetin‐4′‐CH₃, quercetin‐3′‐OCH₃ and quercetin‐3′‐Br are substrates of UGT1A3. Wogonin, baicalein, quercetin‐4′‐Cl, quercetin‐3‐OCH₂OCH₃, quercetin‐3‐O‐arabinoside, quercetin‐4′‐CH₃, quercetin‐3′‐OCH₃ and quercetin‐3′‐Br are the newly reported substrates of UGT1A9. The preferred substrates for UGT1A3 and UGT1A9 contain the hydroxyl group at the C7‐position. The glycon and the position of the B ring have conspicuous influences on the glucuronidation activity, and other chemical structures of flavonoids have minor effects. Conclusions From the quantitative study, UGT1A9 in general has higher glucuronidation efficiency than UGT1A3.     

Raloxifene glucuronidation in liver and intestinal microsomes of humans and monkeys: contribution of UGT1A1, UGT1A8 and UGT1A9

1.?Raloxifene is an antiestrogen that has been marketed for the treatment of osteoporosis, and is metabolized into 6- and 4′-glucuronides by UDP-glucuronosyltransferase (UGT) enzymes. In this study, the in vitro glucuronidation of raloxifene in humans and monkeys was examined using liver and intestinal microsomes and recombinant UGT enzymes (UGT1A1, UGT1A8 and UGT1A9).

2.?Although the K_m and CL_int values for the 6-glucuronidation of liver and intestinal microsomes were similar between humans and monkeys, and species differences in V_max values (liver microsomes, humans?>?monkeys; intestinal microsomes, humans?<?monkeys) were observed, no significant differences were noted in the K_m or S₅₀, V_max and CL_int or CL_max values for the 4′-glucuronidation of liver and intestinal microsomes between humans and monkeys.

3.?The activities of 6-glucuronidation in recombinant UGT enzymes were UGT1A1?>?UGT1A8?>UGT1A9 for humans, and UGT1A8?>?UGT1A1?>?UGT1A9 for monkeys. The activities of 4′-glucuronidation were UGT1A8?>?UGT1A1?>?UGT1A9 in humans and monkeys.

4.?These results demonstrated that the profiles for the hepatic and intestinal glucuronidation of raloxifene by microsomes were moderately different between humans and monkeys.     

Three novel single nucleotide polymorphisms (SNPs) of CYP2S1 gene in Japanese individuals

We analyzed all nine exons and exon-intron junctions of the CYP2S1 gene in 200 Japanese individuals and identified the following three novel single nucleotide polymorphisms (SNPs): 4612G>A (Glu147Glu) in exon 3, 5478C>T (Leu230Leu) and 5479T>G (Leu230Arg, CYP2S1*5A) in exon 5. The allele frequencies were 0.013 for 4612G>A, 0.058 for 5478C>T, and 0.003 for 5479T>G. In addition, a known SNP 1324C>G (Pro74Pro) was detected at a frequency of 0.300.     

染料木素临床不良反应与CYP1A2、UGT1A7基因多态性研究

目的：探讨染料木素临床不良反应与CYP1A2、UGT1A7基因多态性的相关性。方法：114例健康志愿者随机分为试验组与对照组,试验组分别口服染料木素一个剂量50、100、200mg,每人服药1次,观察3d了解有无不良反应发生;用限制性片段长度多态性聚合酶链反应（RFLR-PCR）扩增基因片段并酶切电泳观察分析CYP1A2G2964A、C734A和UGT1A7Trp208Arg的多态性。结果：试验组及对照组的基因型及等位基因分布差异无统计学意义（P〉0.05）。试验组受试者根据有无不良反应的出现分为两组基因,CYP1A2G2964A基因：不良反应组14例受试者中有10例的基因型为G/A（占71.43%）,而无不良反应组41例受试者中有22例的基因型为G/G（占53.66%）;CYP1A2C734A基因：不良反应组13例受试者中有7例的基因型为C/A（占53.85%）,而无不良反应组32例受试者中有16例的基因型为A/A（占50.00%）;UGT1A7Trp208Arg基因：不良反应组15例受试者中有12例的基因型为Trp/Trp（占80.00%）,并且无不良反应组53例受试者中有32例的基因型也为Trp/Trp（占60.38%）。结论：染料木素不良反应组中CYP1A2G2964A基因以G/A型较高,CYP1A2C734A基因以G/G型较高;UGT1A7基因以Trp/Trp型较高。