焦磷酸测序分析ABCG2基因多态性方法的建立

目的:建立ABCG2 34G＞A和421C＞A单核苷酸多态性位点的焦磷酸测序方法.方法:抽取志愿者静脉血入EDTA抗凝管,常规苯酚/氯仿法制备全血gDNA,生物素标记引物对扩增多态位点目的片段,制备生物素标记单链模板,与测序引物退火结合后行焦磷酸测序.分析结果经毛细管电泳测序验证,并进行重复性检验.结果:本文建立了针对ABCG2 34G＞A和421C＞A多态性位点的焦磷酸测序方法,经毛细管电泳测序验证和重复性验证,结果准确可靠.ABCG2 34G和34A等位基因频率分别为79.5％和20.5％;421C和421A等位基因频率分别为72.7％和27.8％,均符合Hardy -Weinberg平衡.结论:本文建立的焦磷酸测序方法可准确、高通量、快速检测ABCG2 34G＞A和421C＞A单核苷酸多态性,并且特别适宜大样本量的临床及科研批量检测需要.     

焦磷酸测序技术检测LAMA5 rs944895基因多态性方法的建立

目的:建立LAMA5 rs944895基因多态性分析的焦磷酸测序方法,并分析该多态性在中国健康人群中的分布频率。方法:制备201例健康人外周血gDNA,应用PyroMark ID焦磷酸测序仪进行LAMA5 rs944895基因多态位点的焦磷酸测序,并经过重复性试验和毛细管电泳测序验证该方法的准确性。结果:建立了LAMA5rs944895基因多态性分析的焦磷酸测序新方法,检出率100%,重复性试验和毛细管电泳测序验证准确可靠。201例健康人中A等位基因频率为72.4%,G为27.6%;AA、AG、GG等位基因型频率分别为53.2%、38.3%和8.5%,符合Hardy-Weinberg平衡(P=0.55)。结论:LAMA5rs944895基因多态性分析的焦磷酸测序新方法具有快速、准确和高通量的优点,适合于在科研和临床应用中推广。     

湖北地区老年人群SLCO1B1基因多态性分析

目的:探讨湖北地区老年人群有机阴离子转运多肽OATP1B1(由SLCO1B1基因编码)单核苷酸多态性(single nucleotide polymorphism, SNP)的分布特征,为老年人群合理用药提供依据。方法:分析1814例湖北地区老年人群样本检测SLCO1B1基因主要突变位点c.388AG及c.521TC的基因型和等位基因(采用PCR-荧光探针法)。结果:SLCO1B1 c.388AG位点等位基因突变发生率为68.96%,c.521TC位点等位基因突变发生率为9.98%;基因型以*1b/*1b及1a/*1b最常见,占69.84%,其次为*1b/*15(11.63%)、*1a/*1a(11.30%)、*1a/*15(4.58%)、*1a/*5(1.54%)、*15/*15(0.94%)、*5/*15(0.17%);不同性别SLCO1B1基因型分布比较无统计学差异(P0.05)。结论:本研究展示了湖北地区老年人群SLCO1B1基因多态性分布,结果对临床调整用药方案具有重要意义。     

SLCO1B1基因多态性与阿托伐他汀的安全性及有效性相关性分析

目的 研究高脂血症患者SLCO1B1基因多态性与阿托伐他汀安全性及有效性的相关性。方法 收集金华市人民医院2017年4月—2018年4月在门诊确诊为高脂血症患者的基本资料,测定纳入患者的SLCO1B1 c.388A>G和c.521T>C的基因多态性,定期随访受试者,并定期测定其甘油三酯、胆固醇、低密度脂蛋白胆固醇及肌酸激酶等相关实验室检查指标。结果 纳入患者SLCO1B1 c.388A>G和c.521T>C等位基因频率分别为72.8%和15.9%。随访期结束后不同基因型患者的血清血脂指标变化率无明显差异。SLCO1B1 c.521T>C基因多态性与阿托伐他汀的安全性有相关性（P=0.005）。结论 SLCO1B1c.388A>G基因多态性对阿托伐他汀降脂疗效及安全性无影响。SLCO1B1 c.521T>C基因多态性与阿托伐他汀的降脂疗效无相关性,但对其安全性有一定影响。     

SLCO1B1和APOE基因多态性对瑞舒伐他汀调脂疗效及不良反应的影响

目的探讨SLCO1B1和APOE基因多态性对瑞舒伐他汀临床疗效及不良反应的影响。方法回顾性分析2016年3月至2017年11月中日友好医院157例原发性高脂血症患者长期口服瑞舒伐他汀调脂,并进行SLCO1B1和APOE基因分型检测的临床资料,统计分析服药前和服药8周后血脂水平、肌酸激酶(CK)水平、肝功能、肾功能、不良反应数据。结果 SLCO1B1388A> G和521T> C的基因突变率分别为58.3%和9.2%;APOE 526C> T和388T> C的基因突变率分别为9.3%和5.7%。服药8周后,SLCO1B1 388A> G和521T> C突变型患者血清低密度脂蛋白(LDL)降低程度高于野生型患者(P=0.047和0.016),APOE526C> T突变型患者血清总胆固醇(TG)降低程度高于野生型患者,组间比较均差异显著(P=0.029)。SLCO1B1388A> G突变型患者与野生型患者相比,血清载脂蛋白A1(apoA1)含量高,apoB含量低,apoA1/apoB比值明显升高(P <0.05)。SLCO1B1 521T> C突变型CK增加人数占比显著多于野生型(89%vs. 68.2%,P <0.05)。SLCO1B1和APOE基因多态性对患者肝、肾功能无显著影响(P> 0.05)。结论 SLCO1B1 388A> G和APOE 526C> T基因多态性影响瑞舒伐他汀的调脂疗效,SLCO1B1521T> C可能与瑞舒伐他汀的不良反应相关。检测SLCO1B1和APOE基因分型有助于规避用药风险。     

SLCO1B1和APOE基因多态性对瑞舒伐他汀疗效及安全性的影响

目的 探讨SLCO1B1和APOE基因多态性对瑞舒伐他汀疗效及安全性的影响。方法 入选2020年10月至2021年10月服用瑞舒伐他汀的冠心病患者300例,根据瑞舒伐他汀服用剂量分为10 mg组及20 mg组,每组150例。检测患者SLCO1B1和APOE基因型,监测患者用药前、用药1、3和6个月后的血脂水平指标,并随访其6个月内肌肉、肝脏和神经系统等不良反应发生情况。结果 SLCO1B1 388A>G、521T>C与APOE 526C>T、388T>C的突变率分别为71.7%、13.8%、7.8%和10.5%。20 mg组中,携带SLCO1B1 521T>C突变或*15突变的患者的肌肉不良反应发生率高于非携带者（P<0.05）,但在10 mg组中差异无统计学意义（P> 0.05）。在全部患者组中,携带APOE 526C>T突变患者总胆固醇（TC）及低密度脂蛋白胆固醇（LDL-C）降低水平显著高于野生型患者（P <0.05）,携带APOE 388T>C突变患者TC及LDL-C降低水平显著低于野生型患者（P <0.05）,...     

1例基因多态性与瑞舒伐他汀疗效个体差异的病例分析

目的 探讨一患者服用瑞舒伐他汀后短期降脂疗效减弱的可能原因。方法 焦磷酸测序技术检测患者ATP结合转运蛋白超家族成员2 （ABCG2）及有机阴离子转运体1B1（SLCO1B1）基因多态性。结果 患者为ABCG2 c.421 CA杂合子,SLCO1B1 c.388GG纯合子,SLCO1B1 c.521TT纯合子。结论 SLCO1B1 c.388G突变有可能与瑞舒伐他汀短期降脂疗效减弱有关。     

SLCO1B1基因T521C多态性对氟伐他汀药代动力学、药效学和药物不良反应影响的系统评价北大核心CSCD

目的研究SLCO1B1基因T521C位点多态性与氟伐他汀药物的药代动力学、药效学和药物不良反应的相关性。方法本文采用计算机检索Cochrane Library、Pub Med、EMBase、Webof Science、中国知网和万方数据库,截至2017年12月。收集有关SLCO1B1基因多态性对氟伐他汀影响的研究。筛选文献并对最终纳入的6篇文献进行质量评价和系统综述。结果 SLCO1B1基因T521C位点多态性对服用氟伐他汀后的低密度脂蛋白浓度和总胆固醇降低有显著影响,对氟伐他汀的体内药代动力学过程没有显著影响,且没有证据证明该位点与氟伐他汀导致肌病的药物不良反应有相关性。结论 SLCO1B1基因T521C位点的多态性显著影响氟伐他汀的药效学,与氟伐他汀的药代动力学及药物不良反应的发生无关。     

ABCB1(2677T>G)、SLCO1B1(521T>C)基因多态性与阿托伐他汀个体化用药的研究

目的:探讨新乡地区人群ABCB1 (2677T>G)和SLCO1B1 (521T>C)单核苷酸多态性(SNPs)对心脑血管疾病患者服用阿托伐他汀的不良反应及降脂疗效的影响。方法:通过荧光原位杂交法、高效液相色谱法等,考察患者SLCO1B1 (521T>C)、ABCB1(2677T>G)SNPs与阿托伐他汀药动学、药效学及不良反应的相关性。结果:在新乡市中心医院的90例入选患者中,SLCO1B1 (521T>C)、ABCB1 (2677T>G)等位基因突变频率分别为18.1%和69.8%。用药前后,SLCO1B1 (521T>C)SNPs与阿托伐他汀的血药浓度有相关性,但对其疗效及肌痛风险影响较小,各基因型间差异无显著性。ABCB1 (2677T>G)各基因型患者阿托伐他汀血药浓度组内差异具有显著性(P<0.05),相比ABCB1 2677T等位基因,携带ABCB1 2677G基因的患者对LDL-C降低作用更强,且出现肝毒性的风险较高。结论:ABCB1 (2677T>G)SNPs与阿托伐他汀的降脂疗效及肝毒性具有相关性,G等位基因可能是阿托伐他汀致肝毒性的因素之一。SLCO1B1 (521T>C)SNPs对阿托伐他汀的降脂疗效及肌痛风险无显著影响。     

冠心病患者SLCO1B1基因多态性分布研究

目的 探讨冠心病(冠状动脉粥样硬化性心脏病)患者SLCO1B1?1b(388 A>G)rs2306283、SLCO1B1?5(521 T>C)rs4149056基因的多态性分布,为临床冠心病患者个体化合理运用他汀类药物提供一定的理论依据.方法 采用横断面研究方法,选取2019年6月至2020年1月就诊于山西医科大学第二...     

Organic anion transporting polypeptide-1B1 haplotypes in Chinese patients

Aim： To detect 388G〉A and 521T〉C variant alleles in the organic anion transporting polypeptide- 1B 1 （OATP 1B 1, encoding gene SLCOIB1） gene. Methods： One hundred and eleven healthy volunteers were screened for OATP1B 1 alleles in our study. PCR-restriction fragment length polymorphism was used to identify the 388G〉A polymorphism and a 1-step tetra-primer method was developed for the determination of 521T〉C mutation. Results： The frequencies of the 388G〉A and 521T〉C variant alleles in the Chinese population were 73.4% and 14.0%, respectively. The frequencies of the SLCO1B1＊ 1b and ＊15 haplotypes were 59.9% and 14.0%, respectively. Condusion： The SLCO1B1＊1b and SLCO1B1＊15 variants are relatively common in the Chinese population. Their frequencies are similar to that in the Japanese, but significantly different from that in Caucasians and blacks.     

Rapid identification of three functionally relevant polymorphisms in the OATP1B1 transporter gene using Pyrosequencing

INTRODUCTION: Clinical evidence suggests there are three single nucleotide polymorphisms (SNPs) of the solute carrier organic anion transporter family member B1 (SLCO1B1) gene for which in vivo evidence for a functional relevance for organic anion transporter polypeptides subgroup C (OATP1B1, formerly OATP-C) has been provided. These genetic variants have been shown to lead to altered pharmacokinetics of OATP1B1 substrates, mainly pravastatin, but also the irinotecan metabolite SN-38, estrone-3-sulfate, and estradiol-17beta-glucuronide. The authors therefore developed reliable and quick screening assays to identify the SLCO1B1 SNPs -11187G>A, 388A>G and 521T>C, in order to facilitate the judgment of their clinical role and to identify allelic frequencies of SNPs and haplotypes in a Caucasian random sample. METHODS: Three simplex Pyrosequencing assays were developed and the three selected SLCO1B1 SNPs were screened for in 250 DNA samples from healthy young female and male unrelated volunteers of Caucasian ethnicity. SLCO1B1 haplotypes involving DNA positions -11187, 388 and 521 were identified by in silico haplotyping. RESULTS: A clear identification of the three single nucleotide polymorphisms in the 250 DNA samples was possible and was verified by routine implementation of 40 control samples obtained by conventional sequencing. The frequencies of the variant alleles -11187A, 388G and 521C were 0.09, 0.47 and 0.12, respectively. All observed frequencies of heterozygous of homozygous carriers of SLCO1B1 alleles were in agreement with the Hardy-Weinberg equilibrium. SLCO1B1 haplotypes reportedly associated with altered substrate pharmacokinetics, i.e., SLCO1B1*15B (-11187G/388G/521C) and *17 (-11187A/388G/521C), were found at allelic frequencies of 0.09 and 0.02, respectively. CONCLUSION: The presently developed Pyrosequencing assays allowed for quick and reliable identification of those SLCO1B1 SNPs that had been proposed to cause functional alternations in OATP1B1 with shown consequences for the pharmacokinetics of drugs that are OATP1B1 substrates.     

Lack of association between SLCO1B1 polymorphism and the lipid-lowering effects of atorvastatin and simvastatin in Chinese individuals

Purpose

There is significant inter-individual variability in the lipid-lowering effects of atorvastatin and simvastatin. Our goal was to investigate the impact of SLCO1B1 genetic polymorphism on the lipid-lowering effects of atorvastatin and simvastatin.

Methods

We recruited 363 unrelated hyperlipidemic patients with the CYP3A4*1/*1, CYP3A5*1/*1, and CYP3AP1*1/*1 genotypes: 189 of these were treated with atorvastatin and 174 were treated with simvastatin as a single-agent therapy (20 mg?day^?1 orally) for 4 weeks. The genotyping of SLCO1B1 c.521T?>?C (p.V174A, OATP-C*5) was performed with allele-specific polymerase chain reaction (AS-PCR), and PCR restriction fragment length polymorphism (RFLP) was performed to detect the carriers of SLCO1B1 c.388A?>?G (p.N130D, OATP-C*1b). Serum triglyceride (TGs), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) levels were determined before and after treatment.

Results

The frequencies of the SLCO1B1 521T?>?C and 388A?>?G variant alleles in Chinese hyperlipidemic patients were found to be 16.2% and 72.1% respectively. After treatment with 20 mg simvastatin or atorvastatin daily for 4 weeks, TC, TG, and LDL-C concentrations were lower than at baseline, on average, by 18.1?±?3.7%, 25.8?±?9.7%, 27.7?±?5.4% in the simvastatin-treated group, and 17.5?±?3.7%, 22.6?±?8.6%, 27.5?±?5.5% in the atorvastatin-treated group respectively, and the mean relative reduction in serum HDL cholesterol did not reach statistical significance (?1.0?±?10.9%, 0.5?±?9.3%). However, no significant differences were observed in the lipid-lowering effects of atorvastatin and simvastatin between subjects with different SLCO1B1 genotypes.

Conclusion

The SLCO1B1 521T?>?C and 388A?>?G variants were found to be relatively common in Chinese patients with essential hyperlipidemia. These frequencies were found to be similar to those observed in healthy Chinese and Japanese individuals, but significantly different from Caucasians and blacks. SLCO1B1 521T?>?C and 388A?>?G polymorphisms may not be associated with the lipid-lowering effects of atorvastatin and simvastatin.     

Frequencies of single nucleotide polymorphisms and haplotypes of organic anion transporting polypeptide 1B1 SLCO1B1 gene in a Finnish population

Objective Organic anion transporting polypeptide 1B1 (OATP1B1) is a drug uptake transporter located at the sinusoidal membrane of hepatocytes. Our aim was to establish high-throughput genotyping assays for the major known single nucleotide polymorphisms (SNP) in the SLCO1B1 gene encoding OATP1B1 and to investigate the frequencies of SNPs and haplotypes of SLCO1B1 in a large Caucasian population.Methods Distribution of SLCO1B1 alleles was determined in 468 healthy Finnish Caucasian subjects at 11 variant sites spanning the entire gene by using allelic discrimination with TaqMan 5′ nuclease assays.Results The allelic frequencies of SLCO1B1 SNPs were 9.7% for g.−11187G>A, 0.4% for g.−11110T>G, 8.0% for g.−10499A>C, 46.2% for c.388A>G, 13.1% for c.411G>A, 13.1% for c.463C>A, 20.2% for c.521T>C, 53.2% for c.571T>C, 46.4% for c.597C>T, 4.0% for c.1929A>C and 48.8% for c.*439T>G. Altogether, 26 haplotypes were statistically inferred. The most common haplotype, with a frequency of 35.6%, contained variant allele C at position c.571, but had the reference nucleotide at all other positions investigated. The functionally significant c.521T>C SNP existed in four major haplotypes, which could be differentiated by the g.−11187G>A, g.−10499A>C and c.388A>G SNPs. The frequencies of these haplotypes were 7.9% for g.−11187G/g.−10499C/c.388G/c.521C (*16), 6.9% for AAGC (*17), 2.7% for GAAC (*5) and 2.4% for GAGC (*15).Conclusion Sequence variations of SLCO1B1 occur at high frequencies in the Caucasian population. Further studies are needed to characterise the effects of SLCO1B1 haplotypes, especially *16, *17, *15 and *5, on drug pharmacokinetics and response, and to determine the frequencies of SLCO1B1 SNPs and haplotypes in different populations.     

Lack of effect of genetic polymorphisms of SLCO1B1 on the lipid-lowering response to pitavastatin in Chinese patients

Aim:

To investigate the SLCO1B1 388A>G and 521T>C polymorphisms in hyperlipidemia patients and evaluate the effect of the two polymorphisms on the lipid-lowering efficacy of pitavastatin.

Methods:

The functional polymorphisms of SLCO1B1 (388A>G and 521T>C) were genotyped in 140 Chinese patients with essential hyperlipidemia using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and one-step tetra-primers ARMS-PCR. Eighty-five patients were enrolled in the clinical trial and given 2 mg of pitavastatin daily for 8 weeks. Total cholesterol (TC), triglyceride (TG), high-density lipoprotein (HDL), and low-density lipoprotein (LDL) serum levels were measured at baseline, after 4 weeks and after 8 weeks of treatment.

Results:

The allele frequencies of SLCO1B1 388A>G and 521T>C in essential hyperlipidemia patients were 71.1% and 11.1%, respectively. The 4- and 8-week treatment with pitavastatin significantly reduced TC, TG, and LDL levels, but there was no statistical difference among patients with wild type, SLCO1B1 388A>G or SLCO1B1 521T>C in the lipid-lowering efficacy of pitavastatin.

Conclusion:

The present study found that the allele frequencies of SLCO1B1 388A>G and 521T>C in Chinese patients with essential hyperlipidemia are comparable to those in healthy Chinese population. SLCO1B1 388A>G and 521T>C do not affect the lipid-lowering efficacy of pitavastatin.     

Global analysis of genetic variation in SLCO1B1

INTRODUCTION: Organic anion transporting polypeptide 1B1 (OATP1B1), encoded by the SLCO1B1 gene, mediates the hepatic uptake of endogenous compounds and xenobiotics, including drugs. The aim of this study was to investigate the diversity of the SLCO1B1 gene at the global level. MATERIALS AND METHODS: Distribution of SLCO1B1 alleles was determined in 941 individuals from 52 populations comprising Africa, the Middle East, Asia, Europe, Oceania and the Americas. DNA samples were genotyped at 12 variant sites spanning the entire gene by TaqMan 5 nuclease allelic discrimination assays. RESULTS: The frequency of the low-activity c.521T>C variant varied markedly between populations. The lowest frequencies were observed in Oceania (0.0%; 95% CI: 0.0-6.4%) and sub-Saharan Africa (1.9%; 95% CI: 0.7-4.8%), and the highest frequencies observed in American native populations (24%; 95% CI: 18-32%) and Europe (18%; 95% CI: 14-23%). Moreover, the c.521C allele (r = 0.505, p < 0.001) and the *1B (c.388G-c.521T; r = -0.405, p = 0.006) and *15 (c.388G-c.521C; r = 0.510, p < 0.001) haplotype frequencies correlated significantly with latitude in the northern hemisphere. Overall, SLCO1B1 genetic distances correlated significantly with geographic distances between populations, assuming likely routes of human migration out of Africa via five waypoints (r = 0.235, p = 0.001). SLCO1B1 diversity was generally far greater within than between populations. CONCLUSION: Functionally significant variants of SLCO1B1 are widely distributed and occur at high frequencies around the world. SLCO1B1 diversity is greater within than between populations, and genetic variation in SLCO1B1 is generally similar to that observed for other autosomal markers. However, selective pressure may have acted on SLCO1B1 during human dispersal favoring low-activity variants in the north.     

Influence of genomic ancestry on the distribution of SLCO1B1, SLCO1B3 and ABCB1 gene polymorphisms among Brazilians

The frequency distribution of SNPs and haplotypes in the ABCB1, SLCO1B1 and SLCO1B3 genes varies largely among continental populations. This variation can lead to biases in pharmacogenetic studies conducted in admixed populations such as those from Brazil and other Latin American countries. The aim of this study was to evaluate the influence of self-reported colour, geographical origin and genomic ancestry on distributions of the ABCB1, SLCO1B1 and SLCO1B3 polymorphisms and derived haplotypes in admixed Brazilian populations. A total of 1039 healthy adults from the north, north-east, south-east and south of Brazil were recruited for this investigation. The c.388A>G (rs2306283), c.463C>A (rs11045819) and c.521T>C (rs4149056) SNPs in the SLCO1B1 gene and c.334T>G (rs4149117) and c.699G>A (rs7311358) SNPs in the SLCO1B3 gene were determined by Taqman 5'-nuclease assays. The ABCB1 c.1236C>T (rs1128503), c.2677G>T/A (rs2032582) and c.3435C>T (rs1045642) polymorphisms were genotyped using a previously described single-base extension/termination method. The results showed that genotype and haplotype distributions are highly variable among populations of the same self-reported colour and geographical region. However, genomic ancestry showed that these associations are better explained by a continuous variable. The influence of ancestry on the distribution of alleles and haplotype frequencies was more evident in variants with large differences in allele frequencies between European and African populations. Design and interpretation of pharmacogenetic studies using these transporter genes should include genomic controls to avoid spurious conclusions based on improper matching of study cohorts from Brazilian populations and other highly admixed populations.