Abstract
RNU2 exists in two functional forms (RNU2-1 and RNU2-2) distinguishable by the presence of a unique 4-bases motif. Detailed investigation of datasets obtained from deep sequencing of five human lung primary tumors revealed that both forms express at a high rate a 19–22nt fragment (miR-U2-1 and -2) from its 3′ region and contains the 4-bases motif. Deep sequencing of independent pools of serum samples from healthy donors and lung cancer patients revealed that miR-U2-1 and -2 are pervasively processed in lung tissue by means of endonucleolytic cleavages and stably exported to the blood. Then, microarrays hybridization experiments of matched normal/tumor samples revealed a significant over-expression of miR-U2-1 in 14 of 18 lung primary tumors. Subsequently, qRT-PCR of miR-U2-1 using serum from 62 lung cancer patients and 96 various controls demonstrated that its expression levels identify lung cancer patients with 79% sensitivity and 80% specificity. miR-U2-1 expression correlated with the presence or absence of lung cancer in patients with chronic obstructive pulmonary disease (COPD), other diseases of the lung – not cancer, and in healthy controls. These data suggest that RNU2-1 is a new bi-functional ncRNA that produces a 19–22nt fragment which may be useful in detecting lung cancer non-invasively in high risk patients.
Highlights
The exploration of the non-protein-coding RNA transcriptome in human cells and tissues has long been focused on profiling microRNAs
Since functional ncRNAs are thought to be protected from degradation by virtue of their association with proteins and packaging into particles, and are stable, Next Generation Sequencing technologies (NGS) offers the unique advantage of being able to detect the simultaneous expression of thousands of functional small RNA transcripts in a single tissue type, revealing a new level of complexity in the production of small ncRNAs in human cells, tissues, and organs under various physiological conditions [2,4,8]
To better understand whether the expression of small ncRNA fragments resulting from alternative processing are associated with the presence of disease, we measured the levels of the fragments produced by the snRNA U2 by several methodologies in the lung tissue and serum of patients with lung cancer, and compared them to the RNU2 levels in the serum of patients at risk for lung cancer as well as normal healthy control subjects
Summary
The exploration of the non-protein-coding RNA (ncRNA) transcriptome in human cells and tissues has long been focused on profiling microRNAs. To better understand whether the expression of small ncRNA fragments resulting from alternative processing are associated with the presence of disease, we measured the levels of the fragments produced by the snRNA U2 by several methodologies in the lung tissue and serum of patients with lung cancer, and compared them to the RNU2 levels in the serum of patients at risk for lung cancer (patients with COPD) as well as normal healthy control subjects.
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