Department of Geriatric Medicine and Nephrology, Osaka University Graduate School of Medicine, Japan.
Atherosclerosis. 2012 Feb;220(2):449-55. doi: 10.1016/j.atherosclerosis.2011.11.017. Epub 2011 Nov 19.
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.
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.
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.
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.
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.
全基因组关联研究（GWAS）已经确定了位于染色体 9p21 位置的导致心血管疾病（CVD）风险的遗传变异。与 CVD 相关的区域毗邻两个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 的最后外显子。目前尚不清楚这些转录本中的哪一个或如何参与动脉粥样硬化的发病机制。
我们评估了以下假设，即 9p21 基因座多态性影响该区域内的转录本（ANRIL、CDKN2A/B）的表达，并且这些转录本通过调节 VSMC 的增殖而导致动脉粥样形成。
我们对横跨感兴趣区域（CDKN2A/B 和 ANRIL，包含 CVD 相关区域）的 18 个 SNP（r（2）<0.8 和 MAF>0.05）进行了基因分型。从 57 名志愿者（69-72 岁）的血液中提取 RNA 和 DNA。对 56 名受试者进行颈动脉超声检查。采用 RT-PCR 测量 CDKN2A/B 和 ANRIL（外显子 1-2 和 17-18）的表达。在用 siRNA 介导的 ANRIL 敲低后，评估培养的 VSMC 中的基因表达和细胞生长。
动脉粥样硬化相关表型的风险等位基因与评估外显子 1-2 时 ANRIL 的表达呈负相关。颈总动脉狭窄与 ANRIL（外显子 1-2）的表达明显降低（P<0.01）相关。在 VSMC 中 ANRIL 的敲低导致 CDKN2A/B 的表达显著变化（P<0.05）和体外细胞生长减少（P<0.05）。
9p21 基因座上与疾病相关的 SNP 主要影响 ANRIL 的表达。总的来说，我们的研究结果表明，9p21 基因座上的几个与 CVD 相关的 SNP 影响 ANRIL 的表达，进而调节细胞生长，可能通过 CDKN2A/B 调节。
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