Wnt5a/Ror2通过激活PKC促进血管平滑肌细胞增殖。

IF 1.7 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY
Yaning Shi, Hongfang Li, Jia Gu, Yongzhen Gong, Xuejiao Xie, Duanfang Liao, Li Qin
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引用次数: 0

摘要

血管平滑肌细胞(VSMCs)增生异常可引起动脉粥样硬化、再狭窄、肺动脉高压等多种血管疾病。然而,Wnt5a对VSMCs增殖的作用及其机制尚不清楚。我们的研究旨在探讨Wnt5a/Ror2是否通过激活蛋白激酶C (PKC)促进血管平滑肌细胞增殖,从而有效缓解血管增生性疾病。材料和方法:采用CCK-8、EdU和平板克隆形成法评价HA-VSMC细胞系的增殖能力。采用标准方法进行Wnt5a基因敲低和过表达。采用共免疫沉淀法研究Wnt5a与Ror2的相互作用。Western blotting和免疫荧光法检测VSMCs中关键蛋白的表达水平。结果:本研究发现,在10%血清刺激的VSMCs增殖过程中,Wnt5a蛋白表达明显增加,且呈时间依赖性。此外,过表达Wnt5a的VSMCs的增殖速度显著加快,而使用siWnt5a敲除Wnt5a则逆转了这种增殖作用。Wnt5a通过与受体酪氨酸激酶样孤儿受体2 (Ror2)结合,上调其表达。进一步研究表明,Wnt5a可诱导VSMCs中PKC的表达,敲低Wnt5a或Ror2可抑制PKC的磷酸化。结论:Wnt5a能有效促进VSMCs的增殖,这可能与Wnt5a与Ror2结合激活PKC有关。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Wnt5a/Ror2 promotes vascular smooth muscle cells proliferation via activating PKC.

Introduction: Abnormal proliferation of vascular smooth muscle cells (VSMCs) can cause various vascular diseases, such as atherosclerosis, restenosis, and pulmonary hypertension. However, the effect and underlying mechanism of Wnt5a on the proliferation of VSMCs remain unclear. Our study aimed to investigate whether Wnt5a/Ror2 promotes vascular smooth muscle cell proliferation via activating protein kinase C (PKC), thereby effectively alleviating vascular proliferative diseases.

Material and methods: The proliferation of HA-VSMC cell line was evaluated by CCK-8, EdU, and Plate clone formation assays. The Wnt5a gene knockdown and overexpression were carried out by standard methods. The interaction between Wnt5a and Ror2 was explored by co-immunoprecipitation. Western blotting and immunofluorescence were used to determine the expression levels of key proteins in VSMCs.

Results: The present study found that the expression of Wnt5a protein increased significantly in the proliferation of VSMCs stimulated by 10% serum in a time-dependent manner. Furthermore, the proliferative rate of VSMCs overexpressing Wnt5a was dramatically accelerated, whereas Wnt5a knockdown using siWnt5a reversed thisproliferative effect. Wnt5a up-regulated the expression of receptor tyrosine kinase-like orphan receptor 2 (Ror2) by binding to it. Further studies indicated that Wnt5a induces the PKC expression in VSMCs and knockdown of Wnt5a or Ror2 could inhibit PKC phosphorylation.

Conclusions: Wnt5a could effectively promote the proliferation of VSMCs, which might be related to the binding of Wnt5a and Ror2 to activate PKC.

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来源期刊
Folia histochemica et cytobiologica
Folia histochemica et cytobiologica 生物-生化与分子生物学
CiteScore
2.80
自引率
6.70%
发文量
56
审稿时长
6-12 weeks
期刊介绍: "Folia Histochemica et Cytobiologica" is an international, English-language journal publishing articles in the areas of histochemistry, cytochemistry and cell & tissue biology. "Folia Histochemica et Cytobiologica" was established in 1963 under the title: ‘Folia Histochemica et Cytochemica’ by the Polish Histochemical and Cytochemical Society as a journal devoted to the rapidly developing fields of histochemistry and cytochemistry. In 1984, the profile of the journal was broadened to accommodate papers dealing with cell and tissue biology, and the title was accordingly changed to "Folia Histochemica et Cytobiologica". "Folia Histochemica et Cytobiologica" is published quarterly, one volume a year, by the Polish Histochemical and Cytochemical Society.
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