Department of Medical Genetics, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China.
PLoS One. 2009 Oct 26;4(10):e7594. doi: 10.1371/journal.pone.0007594.
miRNAs are 17-25 nucleotides long RNA molecules that have been found to regulate gene expression in human cells. There are studies showing that different groups of miRNAs are involved in development of different tissues. In hepatocytes there are reported particular types of miRNAs that regulate gene expression.
We established a human fetal liver cDNA library by a modified cloning protocol. Then plasmid isolation from the colonies was performed. After sequencing and database searching, the miRNAs were recognized. RT-PCR and sequencing were carried out to validate the miRNAs detected. Real-time PCR was used to analyze the expression of each miRNA.
One novel miRNA was discovered, together with another 35 previously-known miRNAs in the fetal liver. Some of them existed in variants. The miRNAs identified were validated by RT-PCR and sequencing. Quantitative analysis showed that they have variable expression.
Our results indicate that a special group of miRNAs may play an important role in fetal liver development in a synergistic manner.
miRNA 是一类长度为 17-25 个核苷酸的 RNA 分子，已经发现它们可以在人类细胞中调节基因表达。有研究表明，不同的 miRNA 组参与了不同组织的发育。在肝细胞中，有报道称存在特定类型的 miRNA 可以调节基因表达。
我们通过改良的克隆方案建立了一个人类胎儿肝脏 cDNA 文库。然后从菌落中提取质粒。测序和数据库搜索后，识别出了 miRNA。通过 RT-PCR 和测序对检测到的 miRNA 进行验证。实时 PCR 用于分析每个 miRNA 的表达。
在胎儿肝脏中发现了一个新的 miRNA，以及另外 35 个已知的 miRNA。其中一些存在变体。通过 RT-PCR 和测序验证了鉴定出的 miRNA。定量分析表明它们的表达存在差异。
我们的结果表明，一组特殊的 miRNA 可能以协同的方式在胎儿肝脏发育中发挥重要作用。
