Allelic Imbalance of mRNA Associated with α2-HS Glycoprotein (Fetuin-A) Polymorphism.
Allelic Imbalance of mRNA Associated with α2-HS Glycoprotein (Fetuin-A) Polymorphism.
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DOI：
10.1155/2015/865053
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发表时间：
2015
期刊：
影响因子：
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通讯作者：
Kakimoto Y
中科院分区：
文献类型：
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作者：
Inaoka Y;Osawa M;Mukasa N;Miyashita K;Satoh F;Kakimoto Y
α2-HS糖蛋白(AHSG)，又称胎球蛋白-A，在人群遗传学上表现出由AHSG∗1和AHSG∗2两个主要等位基因组成的多态。AHSG∗1纯合子的血清水平显著高于AHSG∗2纯合子。本研究探讨了顺式调控表达的分子机制。为了定量杂合子内检测中的等位基因特异性mRNA，建立了RT-PCR方法，该方法使用了两个紧密定位的SNPs中加入的引物。定量测定肝组织和肝培养细胞中AHSG∗1和AHSG∗2的含量分别为2.5倍和6.2倍。观察到两个主要等位基因的mRNA表达差异，这与血清水平的差异是一致的。培养细胞携带rs4917和rs4918的杂合型，而rs2248690的纯合型。这种不平衡不太可能来自启动子位置的SNP。此外，为了研究mRNA降解的影响，细胞培养中的RNA合成被放线菌素-D有效地抑制。两个等位基因之间没有明显的变化。结果表明，这种顺式调控表达的差异可能发生在转录或剪接水平上。
Alpha 2-HS glycoprotein (AHSG), also designated as fetuin-A, exhibits polymorphism in population genetics consisting of two major alleles of AHSG ∗ 1 and AHSG ∗ 2. The serum level in the AHSG ∗ 1 homozygote is significantly higher than that of the AHSG ∗ 2 homozygote. This study examined the molecular mechanism for the cis-regulatory expression. To quantitate allele-specific mRNA in intra-assays of the heterozygote, RT-PCR method employing primers that were incorporated to the two closely located SNPs was developed. The respective magnitudes of AHSG ∗ 1 to AHSG ∗ 2 in the liver tissues and hepatic culture cells of PLC/PRF/5 were determined quantitatively as 2.5-fold and 6.2-fold. The mRNA expressional difference of two major alleles was observed, which is consistent with that in the serum level. The culture cells carried heterozygous genotypes in rs4917 and rs4918, but homozygous one in rs2248690. It was unlikely that the imbalance was derived from the SNP located in the promotor site. Furthermore, to investigate the effect of mRNA degradation, RNA synthesis in the cell culture was inhibited potently by the addition of actinomycin-D. No marked change was apparent between the two alleles. The results indicated that the cis-regulatory expressional difference is expected to occur at the level of transcription or splicing of mRNA.