Sequencing based quantitative and qualitative analyses of the microRNA transcriptome in hepatitis B related hepatocellular carcinoma.

microrna （miRNA）参与了关键的生物学过程，现在很明显，miRNA的改变参与了人类癌症的进展。然而，大量的mirna尚未被发现，这些元素的精确动力学需要被阐明。最近用克隆技术进行的miRNA分析研究表明，测序方法在检测新型miRNA、修饰和精确组成方面具有优势，并且通过克隆计数分析计算的克隆频率具有可重复性。在这里，我们专注于miRNA的测序，以获得与人类肝脏相关的这些转录组的全面概况和表征。在乙型肝炎相关肝细胞癌（HCC）患者中发现新的miRNA和miRNA生物标志物将显示测序的有用性。我们确定了来自HCC的超过314000个mirna和来自邻近正常肝脏（ANL）的超过268000个注册人类mirna的可靠读数。计算生物信息学鉴定了250多个新的mirna。一些新序列来源于已知前体的同一臂，包括miR-21，从而鉴定出三个成熟序列。此外，序列调查揭示了成熟mirna的各种修饰。使用克隆计数分析的表达谱分析用于鉴定在癌症中异常表达的mirna，包括miR-21、miR-122、miR-122*。在这方面，miR-21似乎是HCC的预测性生物标志物。我们相信，测序将加速发现与人类肝癌相关的新型miRNA和miRNA生物标志物。

MicroRNAs (miRNAs) participate in crucial biological processes, and it is now evident that miRNA alterations are involved in the progression of human cancers. A large number of miRNAs, however, have yet to be discovered, and the precise dynamics of these elements need to be elucidated. Recent studies on miRNA profiling performed with cloning techniques suggest that sequencing methods are superior for the detection of novel miRNAs, modifications, and precise compositions, and that cloning frequencies calculated by clone count analysis are reproducible. Here we focus on sequencing of miRNA to obtain a comprehensive profile and characterization of these transcriptomes as they relate to human liver. The usefulness of sequencing will be shown for the discovery of novel miRNAs and miRNA biomarkers in the patients with hepatitis B associated hepatocellular carcinoma (HCC). We identified reliable reads of more than 314000 miRNAs from HCC and more than 268000 from adjacent normal liver (ANL) for registered human miRNAs. Computational bioinformatics identified more than 250 novel miRNAs. Some novel sequences were derived from the same arm of known precursors, including miR-21, resulting in the identification of three mature sequences. Moreover, the sequence survey revealed various modifications of mature miRNAs. Expression profiling using clone count analysis was used to identify miRNAs including miR-21, miR-122, miR-122*, that are expressed aberrantly in cancer. In this regard, miR-21 appears to be a predictive biomarker for HCC. We believe that sequencing will accelerate the discovery of novel miRNAs and miRNA biomarkers involved in human liver cancer.