Ultrasound stimulation induces microRNA expression changes that could be involved in sonication-induced apoptosis
Ryohei Ogawa1, Akihiro Morii2, Akihiko Watanabe2
1Department of Radiological Sciences, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani, Toyama, 930-0194, Japan. ogawa@med.u-toyama.ac.jp.
Purpose:
The purpose of this study is to investigate the involvement of microRNAs (miRNAs) in sonication-induced apoptosis.
Methods:
U937 cells derived from human leukemia were sonicated with 1-MHz ultrasound at 0.4 W/cm(2) and 10 % duty factor for 60 s, a condition inducing apoptosis. The total RNA was extracted from cells at various timings after sonication and subjected to microarray and real-time PCR for miRNA expression analyses.
Results:
Expression of several miRNAs was significantly affected by sonication. For miR-424* and miR-720, whose expressions were eminently decreased by sonication, cell lines overexpressing these miRNAs were established. Conversely, for miR-663B and miR-663, whose expressions were eminently increased by sonication, cell lines inhibiting these miRNA functions were established. When these cell lines were sonicated, a cell line inhibiting miR-663B function significantly increased sonication-induced apoptosis, suggesting this may be involved in cellular responses to sonication. Two genes that could induce apoptosis, KSR2 and CREBZF, were identified as potential target genes of miR-663B since potential target sequences on their 3' UTR mediated to decrease expression of a reporter gene.
Conclusion:
These results suggest that miRNAs may be involved in cellular responses to ultrasound through their expression changes caused by sonication.
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