State Key Laboratory of Oral Disease, West China College of Stomatology, Sichuan University, Chengdu, PR China.
Am J Med Genet A. 2011 Dec;155A(12):2940-8. doi: 10.1002/ajmg.a.34254. Epub 2011 Oct 19.
The noncoding SNP rs7205289, located in the microRNA-140 gene has been associated with cleft palate risk. MiR-140 was found to regulate zebrafish palatal development in vivo and its expression level be reduced by environmental smoke exposure in vitro. Therefore, we sought to investigate whether the A allele of rs7205289 and maternal smoke exposure during the first trimester might contribute to cleft palate risk by regulating microRNA-140. We used in situ hybridization to explore the microRNA-140 expression pattern. A luciferase reporting system and Western blot were used to validate the target of microRNA-140. Mouse palatal mesenchymal cells (MPMC) were transfected with microRNA-140 expression vectors, or treated with cigarette smoke extract. In addition, we performed a hospital-based case-control study in 169 patients with nonsyndromic cleft palate and 306 unaffected controls. We demonstrated microRNA-140 expression in mouse palatal shelves from embryonic days 12 to 15. Pdgfrα was the target of microRNA-140 in MPMC. When these cells were transfected with the minor allele vector or exposed to cigarette smoke extract, they showed a decrease in microRNA-140 expression. Epidemiological analyses showed that infants with CA/AA genotypes and exposed to maternal passive smoking during pregnancy had evidence of synergistic interaction in contributing to cleft palate risk. We concluded that infants with CA/AA genotypes at rs7205289 and maternal passive smoking during the first trimester may synergistically contribute to cleft palate risk by decreasing microRNA-140 during palatal development.
非编码 SNP rs7205289 位于 microRNA-140 基因中，与腭裂风险相关。研究发现，miR-140 可调节体内斑马鱼腭部发育，其表达水平可被体外环境烟雾暴露所降低。因此，我们试图研究 rs7205289 的 A 等位基因和母亲在妊娠早期接触环境烟雾是否通过调节 microRNA-140 导致腭裂风险。我们使用原位杂交技术来探索 microRNA-140 的表达模式。使用荧光素酶报告系统和 Western blot 来验证 microRNA-140 的靶基因。用 microRNA-140 表达载体转染小鼠腭间充质细胞（MPMC），或用香烟烟雾提取物处理。此外，我们在 169 例非综合征性腭裂患者和 306 例无腭裂对照中进行了基于医院的病例对照研究。我们在胚胎第 12 至 15 天的小鼠腭板中证明了 microRNA-140 的表达。Pdgfrα 是 MPMC 中 microRNA-140 的靶基因。当这些细胞转染小等位基因载体或暴露于香烟烟雾提取物时，其 microRNA-140 表达减少。流行病学分析表明，具有 CA/AA 基因型且在妊娠期间暴露于母亲被动吸烟的婴儿有协同作用，增加了腭裂风险。我们得出结论，rs7205289 处携带 CA/AA 基因型且在妊娠早期接触母亲被动吸烟的婴儿可能通过降低腭发育过程中的 microRNA-140 而协同增加腭裂风险。
