PDGFRB promotes dedifferentiation and pulmonary metastasis through rearrangement of cytoskeleton under hypoxic microenvironment in osteosarcoma

IF 4.4 2区生物学 Q2 CELL BIOLOGY

Cellular signalling Pub Date : 2024-11-04 DOI:10.1016/j.cellsig.2024.111501

Enjie Xu , Zhen Huang , Kunpeng Zhu , Jianping Hu , Xiaolong Ma , Yongjie Wang , Jiazhuang Zhu , Chunlin Zhang

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引用次数: 0

Abstract

Background

Osteosarcoma (OS) cells commonly suffer from hypoxia and dedifferentiation, resulting in poor prognosis. We plan to identify the role of hypoxia on dedifferentiation and the associated cellular signaling.

Methods

We performed sphere formation assays and determined spheroid cells as dedifferentiated cells by detecting stem cell-like markers. RNAi assay was used to explore the relationship between hypoxia inducible factor 1 subunit alpha (HIF1A) and platelet derived growth factor receptor beta (PDGFRB). We obtained PDGFRB knockdown and overexpression cells through lentiviral infection experiments and detected the expression of PDGFRB, p-PDGFRB, focal adhesion kinase (FAK), p-FAK, phosphorylated myosin light chain 2 (p-MLC2), and ras homolog family member A (RhoA) in each group. The effects of PDGFRB on cytoskeleton rearrangement and cell adhesion were explored by immunocytochemistry. Wound-healing experiments, transwell assays, and animal trials were employed to investigate the effect of PDGFRB on OS cell metastasis both in vitro and in vivo.

Results

Dedifferentiated OS cells were found to exhibit high expression of HIF1A and PDGFRB, and HIF1A upregulated PDGFRB, subsequently activated RhoA, and increased the phosphorylation of MLC2. PDGFRB also enhanced the phosphorylation of FAK. The OS cell morphology and vinculin distribution were altered by PDGFRB. PDGFRB promoted cell dedifferentiation and had a significant impact on the migration and invasion abilities of OS cells in vitro. In addition, PDGFRB increased pulmonary metastasis of OS cells in vivo.

Conclusion

Our results demonstrated that HIF1A up-regulated PDGFRB under hypoxic conditions, and PDGFRB regulated the actin cytoskeleton, a process likely linked to the activation of RhoA and the phosphorylation of, thereby promoting OS dedifferentiation and pulmonary metastasis.

Abstract Image

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在骨肉瘤缺氧微环境下，PDGFRB通过重排细胞骨架促进去分化和肺转移。

背景：骨肉瘤（Osteosarcoma，OS）细胞普遍存在缺氧和去分化现象，导致预后不良。我们计划确定缺氧对去分化的作用以及相关的细胞信号传导：方法：我们进行了球形形成试验，并通过检测干细胞样标记物确定球形细胞为去分化细胞。我们使用RNAi实验来探讨缺氧诱导因子1亚基α（HIF1A）和血小板衍生生长因子受体β（PDGFRB）之间的关系。我们通过慢病毒感染实验获得了 PDGFRB 敲除和过表达细胞，并检测了各组细胞中 PDGFRB、p-PDGFRB、局灶粘附激酶（FAK）、p-FAK、磷酸化肌球蛋白轻链 2（p-MLC2）和 ras 同源家族成员 A（RhoA）的表达。免疫细胞化学研究了 PDGFRB 对细胞骨架重排和细胞粘附的影响。采用伤口愈合实验、跨孔实验和动物实验来研究PDGFRB对OS细胞体外和体内转移的影响：结果：研究发现，去分化 OS 细胞表现出 HIF1A 和 PDGFRB 的高表达，HIF1A 上调 PDGFRB，随后激活 RhoA，增加 MLC2 的磷酸化。PDGFRB 还增强了 FAK 的磷酸化。PDGFRB改变了OS细胞的形态和Vinculin分布。PDGFRB促进了细胞的去分化，并对体外OS细胞的迁移和侵袭能力产生了显著影响。此外，PDGFRB还能增加OS细胞在体内的肺转移：我们的研究结果表明，在缺氧条件下，HIF1A上调PDGFRB，PDGFRB调节肌动蛋白细胞骨架，这一过程可能与RhoA的激活和磷酸化有关，从而促进了OS的去分化和肺转移。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Cellular signalling 生物-细胞生物学

CiteScore

8.40

自引率

0.00%

发文量

250

审稿时长

27 days

期刊介绍： Cellular Signalling publishes original research describing fundamental and clinical findings on the mechanisms, actions and structural components of cellular signalling systems in vitro and in vivo. Cellular Signalling aims at full length research papers defining signalling systems ranging from microorganisms to cells, tissues and higher organisms.