Abstract
The abnormal expression of microRNAs (miRNAs) is implicated in various human diseases, including cancers. Accordingly, miRNA expressions have been examined in many cancer tissues and blood, but there have been few studies examining smear samples from bone marrow (BM) or peripheral blood. Here we successfully isolated small RNAs from BM smears using a mirVana miRNA Isolation Kit with our original modifications. The isolated small RNAs were then used to measure the levels of representative miRNAs such as miR-155, let-7a, and U6 via real-time PCR with a specific TaqMan probe, although peaks for the ribosomal RNAs (18S, and 28S) were not identified. The PCR curves of the miRNAs were indistinguishable from those from BM living cells from the same donor. Finally, our method for BM smears identified numerous abnormally altered miRNAs (significantly decreased, 39 miRNAs; significantly increased, 27 miRNAs) in follicular lymphomas (FL) compared with normal donors via TaqMan real-time PCR miRNA array. The array indicated that miR-451 showed the greatest decrease in FL (a 345-fold decrease), while miR-338-5p showed the greatest increase in FL (172-fold) relative to normal donors. The miRNAs identified by our study might serve as markers to predict the invasion of FL cells into BM without biopsy. Furthermore, our method will provide a new avenue for the analysis of miRNAs in BM smear samples from various hematologic diseases.Electronic supplementary materialThe online version of this article (doi: 10.1186/2193-1801-3-288) contains supplementary material, which is available to authorized users.
Highlights
MicroRNAs, which are short non-coding singlestranded RNAs of 18–24 mer in length, negatively regulate the target gene expression at the post-transcriptional stage (Bartel 2004; Ambros 2004). miRNAs act to inhibit protein translation or degrade transcripts of the target gene (Bartel 2009)
Patients In the present study, we studied bone marrow (BM) samples from patients with follicular lymphoma (FL) of B celltype who were diagnosed at Hiroshima University Hospital, and from normal donors used as a control
There were no differences in results between the two kits. These findings suggested that, in the BM smears, the RNAs of larger size were completely degraded, whereas the smaller RNAs remained potentially reactive
Summary
MicroRNAs (miRNAs), which are short non-coding singlestranded RNAs of 18–24 mer in length, negatively regulate the target gene expression at the post-transcriptional stage (Bartel 2004; Ambros 2004). miRNAs act to inhibit protein translation or degrade transcripts of the target gene (Bartel 2009). The levels of various miRNAs have been measured in body fluids such as blood (serum or plasma), urine, cerebrospinal fluid, and ascites of patients with cancer (Cortez et al 2011; Etheridge et al 2011). Based on these measurements, some of the circulating miRNAs have. The BM samples are often stored as smears at room temperature (Miura et al 2011). These smears can be transported anywhere while remaining at room temperature, and are convenient for use in diagnoses
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