Genetic variants at the 9p21 locus contribute to atherosclerosis through modulation of ANRIL and CDKN2A/B
Genetic variants at the 9p21 locus contribute to atherosclerosis through modulation of ANRIL and CDKN2A/B
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DOI：
10.1016/j.atherosclerosis.2011.11.017
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发表时间：
2012-02-01
期刊：
影响因子：
5.3
通讯作者：
Rakugi, Hiromi
中科院分区：
文献类型：
--
作者：
Congrains, Ada;Kamide, Kei;Rakugi, Hiromi
全基因组关联研究 (GWAS) 已确定染色体 9p21 位点上导致心血管疾病 (CVD) 风险的遗传变异。 CVD 相关区域邻近两种细胞周期蛋白依赖性激酶抑制剂 (CDKN)2A 和 2B 以及非编码 RNA ANRIL 的最后一个外显子。目前尚不清楚这些转录本中的哪些或如何参与动脉粥样硬化的发病机制。目的：我们评估了以下假设：9p21 位点多态性影响该区域转录本的表达（ANRIL、CDKN2A/B），并且这些转录本通过调节 VSMC 中的增殖促进动脉粥样硬化形成。方法：我们对 18 个 SNP 进行了基因分型 （r(2) < 0.8 且 MAF > 0.05）跨越感兴趣区域：CDKN2A/B 和 ANRIL，涵盖 CVD 相关区域。从 57 名志愿者（69-72 岁）的血液中提取了 RNA 和 DNA。对 56 名受试者进行了颈动脉超声检查。采用 RT-PCR 测量 CDKN2A/B 和 ANRIL（外显子 1-2 和 17-18）表达。在 siRNA 介导的 ANRIL 敲低后，在培养的 VSMC 中评估基因表达和细胞生长。结果：在评估外显子 1-2 时，动脉粥样硬化相关表型的风险等位基因始终与 ANRIL 的较低表达相关。颈总动脉狭窄与 ANRIL（外显子 1-2）表达显着降低（P < 0.01）相关。 VSMC 中 ANRIL 敲除导致体外 CDKN2A/B 表达显着变化（P < 0.05）和细胞生长减少（P < 0.05）。结论：9p21 位点的疾病相关 SNP 主要影响 ANRIL 的表达。总体而言，我们的结果表明 9p21 位点中的几个 CVD 相关 SNP 影响 ANRIL 的表达，而 ANRIL 可能通过 CDKN2A/B 调节进而调节细胞生长。 (C) 2011 Elsevier Ireland Ltd. 保留所有权利。
Genome-wide association studies (GWAS) have identified genetic variants contributing to the risk of cardiovascular disease (CVD) at the chromosome 9p21 locus. The CVD-associated region is adjacent to the two cyclin dependent kinase inhibitors (CDKN)2A and 2B and the last exons of the non-coding RNA, ANRIL. It is still not clear which of or how these transcripts are involved in the pathogenesis of atherosclerosis.Objective: We assessed the hypothesis that 9p21 locus polymorphisms influence the expression of the transcripts in the region (ANRIL, CDKN2A/B) and that these transcripts contribute to atherogenesis through the modulation of proliferation in VSMC.Methods: We genotyped 18 SNPs (r(2) < 0.8 and MAF > 0.05) across the region of interest: CDKN2A/B and ANRIL, encompassing the CVD-associated region. RNA and DNA were extracted from the blood of 57 volunteers (69-72 years old). Carotid ultrasound was performed in 56 subjects. CDKN2A/B and ANRIL (exons 1-2 and 17-18) expression was measured employing RT-PCR. Gene expression and cell growth were evaluated in cultured VSMC after the siRNA-mediated knock-down of ANRIL.Results: The risk alleles for atherosclerosis-related phenotypes were consistently associated with a lower expression of ANRIL when evaluating exons 1-2. Common carotid artery stenosis was associated with a significantly lower (P < 0.01) expression of ANRIL (exons 1-2). ANRIL knock-down in VSMC caused significant variation in expression of CDKN2A/B (P < 0.05) and reduction of cell growth (P < 0.05) in vitro.Conclusion: Disease-associated SNPs at the 9p21 locus predominantly affect the expression of ANRIL. Overall, our results suggest that several CVD-associated SNPs in the 9p21 locus affect the expression of ANRIL, which, in turn modulate cell growth, possibly via CDKN2A/B regulation. (C) 2011 Elsevier Ireland Ltd. All rights reserved.