USP13 Suppresses Colorectal Cancer Angiogenesis by Downregulating VEGFA Expression through Inhibition of the PTEN-AKT Pathway.

IF 4.1 4区医学 Q3 ONCOLOGY

Oncology Research Pub Date : 2025-07-18 eCollection Date: 2025-01-01 DOI:10.32604/or.2025.060440

Guo-Zhi Xu, Han-Yang Guan, Yan-Guan Guo, Yi-Ran Zhang, Jing-Hua Pan, Simin Luo, Hui Ding, Yunlong Pan, Qi Yao

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引用次数: 0

Abstract

Background: Tumor angiogenesis is related to solid tumor occurrence. Ubiquitin-specific peptidase 13 (USP13) is a deubiquitinating enzyme with a pivotal effect on tumor proliferation, metastasis, and tumorigenesis. Nonetheless, its effect on colorectal cancer (CRC) angiogenesis remains poorly understood.

Methods: Human umbilical vein endothelial cells (HUVECs) and CRC cells were cultivated, followed by USP13 knockdown/overexpression using shRNA lentiviral vectors or plasmids. Conditioned media (CM) from treated CRC cells were collected to assess HUVEC migration, invasion, and tube formation. Phosphatase and tensin homolog (PTEN) overexpression and recombinant vascular endothelial growth factor A (VEGFA) rescue experiments were performed. Molecular mechanisms were analyzed via Western blot (PTEN, p-AKT, VEGFA), co-immunoprecipitation (PTEN ubiquitination), and in vivo nude mice study to detect the role of USP13 in tumor angiogenesis.

Results: USP13 expression in CRC cells is downregulated and negatively related to platelet endothelial cell adhesion molecule-1 (CD31) expression. Furthermore, the conditioned medium from CRC cells with USP13 knockdown significantly promoted HUVEC migration, invasion, and tube formation, while USP13 overexpression exerted the opposite effect. Additionally, USP13 overexpression significantly increased PTEN expression while decreasing protein kinase B (AKT) phosphorylation levels. Concurrently, USP13 overexpression significantly reduced PTEN ubiquitination, whereas USP13 knockdown remarkably increased this modification. Overexpression of PTEN in sh-USP13 CRC cells decreased the expression levels of VEGFA and p-AKT. USP13 also inhibited tumor angiogenesis through downregulating VEGFA, and recombinant VEGFA blocked the inhibition of the conditioned medium from USP13-overexpressing CRC cells against HUVEC angiogenesis in vivo.

Conclusions: USP13 downregulated VEGFA and inhibited tumor angiogenesis via the PTEN-AKT pathway.

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USP13通过抑制PTEN-AKT通路下调VEGFA表达抑制结直肠癌血管生成。

背景：肿瘤血管生成与实体瘤的发生有关。泛素特异性肽酶13 （USP13）是一种去泛素化酶，在肿瘤增殖、转移和肿瘤发生中起关键作用。尽管如此，其对结直肠癌（CRC）血管生成的影响仍然知之甚少。方法：培养人脐静脉内皮细胞（HUVECs）和结直肠癌细胞，利用shRNA慢病毒载体或质粒敲低/过表达USP13。从处理过的结直肠癌细胞中收集条件培养基（CM）来评估HUVEC的迁移、侵袭和管形成。进行了磷酸酶和紧张素同源物（PTEN）过表达和重组血管内皮生长因子A （VEGFA）的抢救实验。通过Western blot （PTEN、p-AKT、VEGFA）、共免疫沉淀（PTEN泛素化）和体内裸鼠实验分析USP13在肿瘤血管生成中的分子机制。结果：USP13在结直肠癌细胞中表达下调，并与血小板内皮细胞粘附分子-1（血小板内皮细胞粘附分子-1，CD31）表达负相关。此外，USP13敲低的CRC细胞条件培养基显著促进HUVEC的迁移、侵袭和小管形成，而USP13过表达则起到相反的作用。此外，USP13过表达显著增加PTEN表达，同时降低蛋白激酶B （AKT）磷酸化水平。同时，USP13过表达显著降低PTEN泛素化，而USP13敲低显著增加PTEN泛素化。sh-USP13 CRC细胞中PTEN的过表达降低了VEGFA和p-AKT的表达水平。USP13也通过下调VEGFA抑制肿瘤血管生成，重组VEGFA阻断了USP13过表达CRC细胞条件培养基对HUVEC血管生成的抑制作用。结论：USP13通过PTEN-AKT通路下调VEGFA，抑制肿瘤血管生成。

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

Oncology Research 医学-肿瘤学

CiteScore

4.40

自引率

0.00%

发文量

审稿时长

3 months

期刊介绍： Oncology Research Featuring Preclinical and Clincal Cancer Therapeutics publishes research of the highest quality that contributes to an understanding of cancer in areas of molecular biology, cell biology, biochemistry, biophysics, genetics, biology, endocrinology, and immunology, as well as studies on the mechanism of action of carcinogens and therapeutic agents, reports dealing with cancer prevention and epidemiology, and clinical trials delineating effective new therapeutic regimens.