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低强度超声对小鼠 ST2 骨髓基质细胞的遗传应答。

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

机构信息

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.

出版信息

Mol Med Rep. 2021 Mar;23(3). doi: 10.3892/mmr.2020.11812. Epub 2021 Jan 5.

DOI:10.3892/mmr.2020.11812
PMID:33398373
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7821223/
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, 25 mW/cm 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 or the heat shock proteins was associated mainly with the biological functions of bone physiology and protein folding or apoptosis, respectively. Gene network T containing transcription factors 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.

摘要

尽管低强度超声(LIUS)是一种临床确立的程序,但 LIUS 在遗传水平上对早期细胞的影响尚未得到研究。本研究使用全基因组微阵列和计算基因表达分析工具,研究了 LIUS 对骨髓基质细胞(BMSCs)的早期反应基因。将小鼠 ST2 BMSCs 用 LIUS [I,25 mW/cm2,频率为 1.11 MHz 的脉冲波模式(0.2-s 爆发正弦波以 1 kHz 的频率重复)处理 20 min] 处理,然后在 37°C 下培养 0.5、1 和 3 h。使用 GeneChip 高密度寡核苷酸微阵列和 Ingenuity 通路分析工具评估基因表达变化的时间过程。结果通过逆转录-定量聚合酶链反应(RT-qPCR)进行验证。单次 LIUS 照射不会影响细胞形态、细胞生长或碱性磷酸酶活性。然而,LIUS 处理后 0.5 至 3 h 之间,有 61 个上调基因和 103 个下调基因被鉴定出来。从上调基因中鉴定出两个显著的基因网络,分别标记为 E 和 H,而从下调基因中鉴定出第三个网络,标记为 T。包含即刻早期基因 或热休克蛋白 的基因网络 E 或 H 主要与骨生理学和蛋白质折叠或细胞凋亡的生物学功能相关。包含转录因子 的基因网络 T 也与基因表达的生物学功能相关。RT-qPCR 表明,基因网络 E 和 H 中的几个基因的表达在 LIUS 处理的细胞中升高。LIUS 被证明在短时间应用于小鼠 ST2 BMSCs 后能诱导基因表达。本研究的结果为阐明 LIUS 对细胞影响的详细分子机制提供了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d02/7821223/ab37f67bffd8/mmr-23-03-11812-g07.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d02/7821223/2e617afd1891/mmr-23-03-11812-g00.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d02/7821223/e99614034022/mmr-23-03-11812-g02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d02/7821223/7e9eac51a260/mmr-23-03-11812-g03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d02/7821223/72190618491d/mmr-23-03-11812-g04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d02/7821223/7af6970e1ca6/mmr-23-03-11812-g05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d02/7821223/432bee4af9b5/mmr-23-03-11812-g06.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d02/7821223/ab37f67bffd8/mmr-23-03-11812-g07.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d02/7821223/2e617afd1891/mmr-23-03-11812-g00.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d02/7821223/d0df72b22654/mmr-23-03-11812-g01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d02/7821223/e99614034022/mmr-23-03-11812-g02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d02/7821223/7e9eac51a260/mmr-23-03-11812-g03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d02/7821223/72190618491d/mmr-23-03-11812-g04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d02/7821223/7af6970e1ca6/mmr-23-03-11812-g05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d02/7821223/432bee4af9b5/mmr-23-03-11812-g06.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d02/7821223/ab37f67bffd8/mmr-23-03-11812-g07.jpg

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