Department of Animal Science, North Carolina State University, Raleigh, NC 27695-7621, USA.
Anim Genet. 2010 Aug;41(4):357-64. doi: 10.1111/j.1365-2052.2009.02015.x. Epub 2010 Jan 7.
MicroRNA (miRNA) is a family of small regulatory RNAs that post-transcriptionally regulate many biological functions including growth and development. Recently, the expression of chicken miRNA miR-143 was identified by using a deep sequencing approach. In other vertebrate species, miR-143 functions as a regulator of adipocyte differentiation and as a tumour suppressor. However, little is known about the biological function(s) of miR-143 in chickens. To study the functions of chicken miR-143, DNA microarray analysis and a dual luciferase reporter assay were employed to identify genes directly targeted by miR-143 as well as other biologically relevant genes. Microarray analysis indicated that 124 genes were differentially expressed upon in vitro anti-miR-143 treatment in embryonic chick splenocytes (P-value cutoff <0.01). Many of these genes are associated with cell proliferation, apoptosis and tumourigenesis. Six of the up-regulated genes possess at least one potential miR-143 binding site in their 3'UTRs, of these the binding sites of PYCR2, PSTPIP1 and PDCD5 were validated by an in vitro luciferase reporter assay. In addition, several potential targets with important biological functions were identified by the miRanda algorithm and experimentally confirmed. These targets include KLF5, MAP3K7, TARDBP and UBE2E3, which have conserved miR-143 binding sites across multiple vertebrate species. Potential chicken specific miR-143 target sites were also validated for LPIN1, PCK2, PYCR2, METTL14, SLC2A2 and TNFSF10. Overall, the current study suggests that miR-143 is ubiquitously expressed among tissues and is likely to be involved in the regulation of cell proliferation and apoptosis.
微小 RNA (miRNA) 是一类小的调节 RNA，可在后转录水平上调节许多生物学功能，包括生长和发育。最近，使用深度测序方法鉴定了鸡 miRNA miR-143 的表达。在其他脊椎动物物种中，miR-143 作为脂肪细胞分化的调节剂和肿瘤抑制因子发挥作用。然而，关于 miR-143 在鸡中的生物学功能知之甚少。为了研究鸡 miR-143 的功能，采用 DNA 微阵列分析和双荧光素酶报告基因检测法来鉴定 miR-143 直接靶向的基因和其他具有生物学相关性的基因。微阵列分析表明，在胚胎鸡脾细胞体外抗 miR-143 处理后，有 124 个基因表达差异（P 值截止值 <0.01）。这些基因中的许多与细胞增殖、凋亡和肿瘤发生有关。上调基因中有 6 个在其 3'UTR 中至少有一个潜在的 miR-143 结合位点，其中 PYCR2、PSTPIP1 和 PDCD5 的结合位点通过体外荧光素酶报告基因检测得到验证。此外，通过 miRanda 算法和实验验证，还鉴定了几个具有重要生物学功能的潜在靶标。这些靶标包括 KLF5、MAP3K7、TARDBP 和 UBE2E3，它们在多种脊椎动物物种中具有保守的 miR-143 结合位点。还验证了 LPIN1、PCK2、PYCR2、METTL14、SLC2A2 和 TNFSF10 的潜在鸡特异性 miR-143 靶位点。总的来说，本研究表明 miR-143 在组织中广泛表达，可能参与细胞增殖和凋亡的调节。
