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0.5 戈瑞 X 射线照射诱导细胞骨架重排和成骨细胞分化。

0.5‑Gy X‑ray irradiation induces reorganization of cytoskeleton and differentiation of osteoblasts.

机构信息

Department of Orthopedics, The Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu 215004, P.R. China.

出版信息

Mol Med Rep. 2021 May;23(5). doi: 10.3892/mmr.2021.12018. Epub 2021 Mar 24.

DOI:10.3892/mmr.2021.12018
PMID:33760136
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7986016/
Abstract

Osteoblasts are sensitive to ionizing radiation. The small GTPase RhoA and its effector Rho‑associated protein kinase (ROCK) are critical to several cellular functions, including cytoskeleton reorganization, cell survival, and cell differentiation. However, whether the RhoA/ROCK signaling pathway is involved in the regulation of osteoblast cytoskeleton reorganization and differentiation induced by low‑dose X‑ray irradiation remains to be determined. The aim of the present study was to investigate the role of the RhoA/ROCK signaling pathway in mediating differentiation of osteoblasts and reorganization of the cytoskeleton under low‑dose X‑ray irradiation. Osteoblasts were pretreated with the ROCK kinase‑specific inhibitor (Y‑27632) before exposure to low‑dose X‑ray irradiation. The changes of F‑actin in MC3T3 cells were observed at different time points following X‑ray irradiation. Cell Counting Kit‑8 assay, alkaline phosphatase activity, Alizarin red staining and western blotting were used to detect the proliferation and differentiation of osteoblasts after 0.5‑Gy X‑ray irradiation. In the present study, low‑dose X‑ray irradiation promoted the expression of genes associated with the cytoskeleton reorganization. Indeed, the results showed that, 0.5‑Gy X‑ray irradiation can induce reorganization of cytoskeleton and promote differentiation of osteoblasts through the RhoA/ROCK signaling pathway. Additionally, inhibiting ROCK activity blocked low‑dose X‑ray irradiation‑induced LIMK2 phosphorylation, stress fiber formation and cell differentiation. Thus, these results demonstrated the excitatory effects of low‑dose X‑ray irradiation on MC3T3‑E1 cells, including reorganization of the cytoskeleton and differentiation of osteoblasts.

摘要

成骨细胞对电离辐射敏感。小分子 GTP 酶 RhoA 及其效应物 Rho 相关蛋白激酶（ROCK）对于几种细胞功能至关重要，包括细胞骨架重排、细胞存活和细胞分化。然而，RhoA/ROCK 信号通路是否参与调节低剂量 X 射线照射诱导的成骨细胞细胞骨架重排和分化仍有待确定。本研究旨在探讨 RhoA/ROCK 信号通路在介导低剂量 X 射线照射诱导的成骨细胞分化和细胞骨架重排中的作用。在暴露于低剂量 X 射线照射之前，用 ROCK 激酶特异性抑制剂（Y-27632）预处理成骨细胞。在 X 射线照射后不同时间点观察 MC3T3 细胞中 F-肌动蛋白的变化。使用细胞计数试剂盒-8 测定法、碱性磷酸酶活性、茜素红染色和蛋白质印迹法检测 0.5Gy X 射线照射后成骨细胞的增殖和分化。在本研究中，低剂量 X 射线照射促进了与细胞骨架重排相关的基因的表达。事实上，结果表明，0.5Gy X 射线照射可通过 RhoA/ROCK 信号通路诱导细胞骨架重排并促进成骨细胞分化。此外，抑制 ROCK 活性阻断了低剂量 X 射线照射诱导的 LIMK2 磷酸化、应力纤维形成和细胞分化。因此，这些结果表明低剂量 X 射线照射对 MC3T3-E1 细胞具有兴奋作用，包括细胞骨架重排和成骨细胞分化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/132d/7986016/16f6b02fd2d0/mmr-23-05-12018-g10.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/132d/7986016/d39066b14b62/mmr-23-05-12018-g09.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/132d/7986016/16f6b02fd2d0/mmr-23-05-12018-g10.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/132d/7986016/e8979b5db781/mmr-23-05-12018-g00.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/132d/7986016/ca1bde9c356a/mmr-23-05-12018-g01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/132d/7986016/d05357b16c20/mmr-23-05-12018-g02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/132d/7986016/7a7c9752ad44/mmr-23-05-12018-g03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/132d/7986016/0eacd7d1db99/mmr-23-05-12018-g04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/132d/7986016/f9dfb00c482c/mmr-23-05-12018-g05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/132d/7986016/5d312f16229b/mmr-23-05-12018-g06.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/132d/7986016/7943679db08d/mmr-23-05-12018-g07.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/132d/7986016/d7d827325f38/mmr-23-05-12018-g08.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/132d/7986016/d39066b14b62/mmr-23-05-12018-g09.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/132d/7986016/16f6b02fd2d0/mmr-23-05-12018-g10.jpg

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