MDR1基因表达与调控的研究
发布时间：2019-05-11 22:02
【摘要】： 目的:①探讨汉族人群中多药耐药基因1(multidrug resistance gene 1,MDR1) C3435T位点多态性对外周血单个核细胞(peripheral blood mononuclear cells,PBMCs)中mRNA表达的影响。②通过研究苯巴比妥(phenobarbitone,PB)对PBMCs MDR1表达的影响,了解PBMCs MDR1的表达是否也能被诱导,并研究了PBMCs中MDR1与孕烷X受体(pregnane X receptor,PXR)表达的关系,初步探讨其意义。 方法:①以163名(男性98名,女性65名)无亲缘关系的中国健康汉族儿童的外周血标本为研究对象,采用PCR-RFLP技术检测其MDR1基因C3435T位点的基因型,实时荧光定量PCR法检测其PBMCs MDR1 mRNA表达水平。②将36名健康儿童的PBMCs分离后,将其分编为无培养组(不进行培养和药物处理)、自然培养组(只单纯培养不进行药物处理)及PB培养组(加入终浓度为40μm/L苯巴比妥处理)。将自然培养组与PB培养组培养24h后,实时荧光定量PCR检测这三组细胞的MDR1及PXR mRNA表达水平。③Hardy-Weinberg检验采用卡方检验。不同分组间的PBMCs MDR1或PXR表达水平的比较采用t检验或单因素方差分析,MDR1及PXR的表达水平相关性分析使用Spearman法。P0.05表示差异有统计学意义。 结果:①在163名健康中国汉族儿童中,63名为野生型纯合子3435CC型,78名为杂合子CT型,22名为突变型纯合子TT型,CC、CT与TT各基因型的发生频率分别为38.7%、47.9%与13.5%;其C等位基因频率为62.6%,T等位基因频率为37.4%;C3435T多态性位点的基因型频率分布符合Hardy-Weinberg平衡(P0.05);CC、CT和TT各基因型组间PBMCs MDR1 mRNA表达水平分别为(4.562±3.383)×10-3、(4.673±3.710)×10-3和(4.489±2.928)×10-3,差异无统计学意义(P0.05)。②未培养组、自发培养组和PB培养组MDR1 mRNA表达水平分别为(4.475±2.980)×10-3、(4.991±4.165)×10-3及(7.265±5.067)×10-3 , PXR mRNA在以上三组中的表达水平分别为(2.073±1.335)×10-3、(1.836±1.467)×10-3及(3.014±2.238)×10-3。PB培养组MDR1和PXR表达水平均高于未培养组和自发培养组,差异有统计学意义(P0.05),而未培养组和自发培养组之间的MDR1和PXR表达水平差异无统计学意义(P0.05);相关性分析提示,苯巴比妥处理后MDR1及PXR mRNA的表达水平呈正相关关系(r=0.517,P=0.0012)。 结论:①汉族人群中MDR1基因C3435T多态性对PBMCs mRNA表达并没有显著性影响,对于汉族儿童C3435T位点多态性不能作为推测MDR1表达水平的依据。②苯巴比妥可诱导PBMCs MDR1表达,提示PBMCs MDR1表达也能被诱导。苯巴比妥诱导后PXR与MDR1的表达水平呈正相关关系,提示PXR可能参与了PBMCs MDR1基因的表达调控。
[Abstract]:Objective: 1 to investigate the effect of multidrug resistance gene 1 (multidrug resistance gene 1, MDR1) C3435T polymorphism in peripheral blood mononuclear cells (peripheral blood mononuclear cells,PBMCs) on the expression of mRNA in peripheral blood mononuclear cells (peripheral blood mononuclear cells,PBMCs) in Han population. (2) the expression of phenobarbital (phenobarbitone,) was studied by studying the expression of MDR1 in peripheral blood mononuclear cells (PBMCs). The effect of PB) on the expression of PBMCs MDR1 was investigated to see if the expression of PBMCs MDR1 could also be induced, and the relationship between the expression of MDR1 and progesterone X receptor (pregnane X receptor,PXR in PBMCs was studied, and its significance was discussed. Methods: 1 the peripheral blood samples of 163 unrelated Chinese healthy Han children (98 males and 65 females) were used to detect the genotype of C3435T locus of MDR1 gene by PCR-RFLP. The expression of PBMCs MDR1 mRNA was detected by real-time fluorescence quantitative PCR. (2) 36 healthy children were divided into non-culture group (no culture and drug treatment) after PBMCs was isolated and divided into non-culture group (no culture and drug treatment). Natural culture group (simple culture without drug treatment) and PB culture group (with final concentration of 40 渭 m / L phenobarbital). The expression levels of MDR1 and PXR mRNA were detected by real-time fluorescence quantitative PCR after culture in natural culture group and PB culture group for 24 hours. The 3Hardy-Weinberg test was performed by chi-square test. T test or single factor variance analysis were used to compare the expression levels of PBMCs MDR1 or PXR among different groups. Spearman method was used to analyze the correlation between the expression levels of MDR1 and PXR. P 0.05 showed that the difference was statistically significant. Results: 1among 163 healthy Chinese Han children, 63 were wild type homozygote 3435CC, 78 were heterozygote CT and 22 were mutant homozygote TT. The frequency of CC,CT and TT was 38.7%, respectively. 47.9% and 13.5%; The frequency of C allele and T allele were 62.6% and 37.4%, respectively. The genotype frequencies of C allele and T allele were in accordance with Hardy-Weinberg equilibrium (P0.05). The expression levels of PBMCs MDR1 mRNA in CC,CT and TT genotypic groups were (4.562 卤3.383) 脳 10 鈮，
本文编号：2474885
[Abstract]:Objective: 1 to investigate the effect of multidrug resistance gene 1 (multidrug resistance gene 1, MDR1) C3435T polymorphism in peripheral blood mononuclear cells (peripheral blood mononuclear cells,PBMCs) on the expression of mRNA in peripheral blood mononuclear cells (peripheral blood mononuclear cells,PBMCs) in Han population. (2) the expression of phenobarbital (phenobarbitone,) was studied by studying the expression of MDR1 in peripheral blood mononuclear cells (PBMCs). The effect of PB) on the expression of PBMCs MDR1 was investigated to see if the expression of PBMCs MDR1 could also be induced, and the relationship between the expression of MDR1 and progesterone X receptor (pregnane X receptor,PXR in PBMCs was studied, and its significance was discussed. Methods: 1 the peripheral blood samples of 163 unrelated Chinese healthy Han children (98 males and 65 females) were used to detect the genotype of C3435T locus of MDR1 gene by PCR-RFLP. The expression of PBMCs MDR1 mRNA was detected by real-time fluorescence quantitative PCR. (2) 36 healthy children were divided into non-culture group (no culture and drug treatment) after PBMCs was isolated and divided into non-culture group (no culture and drug treatment). Natural culture group (simple culture without drug treatment) and PB culture group (with final concentration of 40 渭 m / L phenobarbital). The expression levels of MDR1 and PXR mRNA were detected by real-time fluorescence quantitative PCR after culture in natural culture group and PB culture group for 24 hours. The 3Hardy-Weinberg test was performed by chi-square test. T test or single factor variance analysis were used to compare the expression levels of PBMCs MDR1 or PXR among different groups. Spearman method was used to analyze the correlation between the expression levels of MDR1 and PXR. P 0.05 showed that the difference was statistically significant. Results: 1among 163 healthy Chinese Han children, 63 were wild type homozygote 3435CC, 78 were heterozygote CT and 22 were mutant homozygote TT. The frequency of CC,CT and TT was 38.7%, respectively. 47.9% and 13.5%; The frequency of C allele and T allele were 62.6% and 37.4%, respectively. The genotype frequencies of C allele and T allele were in accordance with Hardy-Weinberg equilibrium (P0.05). The expression levels of PBMCs MDR1 mRNA in CC,CT and TT genotypic groups were (4.562 卤3.383) 脳 10 鈮，
本文编号：2474885
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