Genetic response to low‑intensity ultrasound on mouse ST2 bone marrow stromal cells

Tabuchi,Yoshiaki; Hasegawa,Hideyuki; Suzuki,Nobuo; Furusawa,Yukihiro; Hirano,Tetsushi; Nagaoka,Ryo; Hirayama,Jun; Hoshi,Nobuhiko; Mochizuki,Takashi

doi:10.3892/mmr.2020.11812

Genetic response to low‑intensity ultrasound on mouse ST2 bone marrow stromal cells

Authors:
- Yoshiaki Tabuchi
- Hideyuki Hasegawa
- Nobuo Suzuki
- Yukihiro Furusawa
- Tetsushi Hirano
- Ryo Nagaoka
- Jun Hirayama
- Nobuhiko Hoshi
- Takashi Mochizuki
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Affiliations: Division of Molecular Genetics Research, Life Science Research Center, University of Toyama, Toyama 930-0194, Japan, Graduate School of Science and Engineering, University of Toyama, Toyama 930‑8555, Japan, Noto Marine Laboratory, Institute of Nature and Environmental Technology, Kanazawa University, Ishikawa 927‑0553, Japan, Department of Liberal Arts and Sciences, Toyama Prefectural University, Toyama 939-0398, Japan, Department of Clinical Engineering, Faculty of Health Sciences, Komatsu University, Komatsu 923‑0961, Japan, Laboratory of Animal Molecular Morphology, Department of Animal Science, Graduate School of Agricultural Science, Kobe University, Kobe 657‑8501, Japan, Medical Ultrasound Laboratory, Co., Ltd., Tokyo 250‑0014, Japan
Published online on: December 23, 2020 https://doi.org/10.3892/mmr.2020.11812
Article Number: 173
Copyright: © Tabuchi et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Although low‑intensity ultrasound (LIUS) is a clinically established procedure, the early cellular effect of LIUS on a genetic level has not yet been studied. The current study investigated the early response genes elicited by LIUS in bone marrow stromal cells (BMSCs) using global‑scale microarrays and computational gene expression analysis tools. Mouse ST2 BMSCs were treated with LIUS [I_SATA, 25 mW/cm² for 20 min with a frequency of 1.11 MHz in a pulsed‑wave mode (0.2‑s burst sine waves repeated at 1 kHz)], then cultured for 0.5, 1 and 3 h at 37˚C. The time course of changes in gene expression was evaluated using GeneChip^® high‑density oligonucleotide microarrays and Ingenuity^® Pathway Analysis tools. The results were verified by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). A single exposure of LIUS did not affect cell morphology, cell growth or alkaline phosphatase activity. However, 61 upregulated and 103 downregulated genes were identified from 0.5 to 3 h after LIUS treatment. Two significant gene networks, labeled E and H, were identified from the upregulated genes, while a third network, labeled T, was identified from the downregulated genes. Gene network E or H containing the immediate‑early genes FBJ osteosarcoma oncogene and early growth response 1 or the heat shock proteins heat shock protein 1a/b was associated mainly with the biological functions of bone physiology and protein folding or apoptosis, respectively. Gene network T containing transcription factors fos‑like antigen 1 and serum response factor was also associated with the biological functions of the gene expression. RT‑qPCR indicated that the expression of several genes in the gene networks E and H were elevated in LIUS‑treated cells. LIUS was demonstrated to induce gene expression after short application in mouse ST2 BMSCs. The results of the present study provide a basis for the elucidation of the detailed molecular mechanisms underlying the cellular effects of LIUS.

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