Phosphorylation of RelA/p65 Ser536 inhibits the progression and metastasis of hepatocellular carcinoma by mediating cytoplasmic retention of NF-κB p65.

IF 3.8 3区 医学 Q2 GASTROENTEROLOGY & HEPATOLOGY
Gastroenterology Report Pub Date : 2024-11-04 eCollection Date: 2024-01-01 DOI:10.1093/gastro/goae094
Wentao Zuo, Haoyang Ma, Jianghui Bi, Tiaolan Li, Yifeng Mo, Shiyu Yu, Jia Wang, Beiqing Li, Jinfeng Huang, Yongwen Li, Li Li
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引用次数: 0

Abstract

Background: Intrahepatic and extrahepatic metastases contribute to the high recurrence rate and mortality of hepatocellular carcinoma (HCC). Constitutive activation of nuclear factor-κB (NF-κB) is a crucial feature of HCC. NF-κB p65 (p50-p65) is the most common dimeric form. Ser536 acts as an essential phosphorylation site of RelA/p65. However, the effect of RelA/p65 Ser536 phosphorylation on progression and metastases during intermediate and advanced HCC has not been reported.

Methods: Phosphorylation of RelA/p65 (p-p65 Ser536) and NF-κB p65 were detected by using immunohistochemical staining in HCC tissue samples. The biological effects of RelA/p65 Ser536 phosphorylation were evaluated by using xenograft and metastasis models. NF-κB p65 nuclear translocation was detected by using Western blotting. The binding of NF-κB p65 to the BCL2, SNAIL, and MMP9 promoters was detected by using chromatin immunoprecipitation. The biological effects on proliferation, migration, invasion, and epithelial-mesenchymal transition were assessed by using tetrazolium-based colorimetry, colony formation, EdU incorporation, flow cytometry, cell wound healing, and transwell assay.

Results: NF-κB p65 is highly expressed, while p-p65 Ser536 is not well expressed in intermediate and advanced HCC tissues. In vivo experiments demonstrated that a phosphorylation-mimetic mutant of RelA/p65 Ser536 (p65/S536D) prevents tumor progression and metastasis. In vitro experiments showed that p65/S536D inhibits proliferation, migration, and invasion. Mechanistically, RelA/p65 Ser536 phosphorylation inhibits NF-κB p65 nuclear translocation and reduces NF-κB p65 binding to the BCL2, SNAIL, and MMP9 promoters.

Conclusions: RelA/p65 Ser536 phosphorylation was detrimental to NF-κB p65 entry into the nucleus and inhibited HCC progression and metastasis by reducing BCL2, SNAIL, and MMP9. The phosphorylation site of RelA/p65 Ser536 has excellent potential to be a promising target for NF-κB-targeted therapy in HCC.

RelA/p65 Ser536的磷酸化通过介导NF-κB p65的细胞质滞留抑制肝细胞癌的进展和转移。
背景:肝内和肝外转移是肝细胞癌(HCC)复发率和死亡率居高不下的原因之一。核因子-κB(NF-κB)的持续激活是 HCC 的一个重要特征。NF-κB p65(p50-p65)是最常见的二聚体形式。Ser536 是 RelA/p65 的重要磷酸化位点。然而,RelA/p65 Ser536磷酸化对中晚期HCC进展和转移的影响尚未见报道:方法:采用免疫组化染色法检测 HCC 组织样本中 RelA/p65 (p-p65 Ser536)和 NF-κB p65 的磷酸化情况。利用异种移植和转移模型评估了 RelA/p65 Ser536 磷酸化的生物学效应。采用 Western 印迹法检测了 NF-κB p65 的核转位。染色质免疫沉淀法检测了 NF-κB p65 与 BCL2、SNAIL 和 MMP9 启动子的结合。利用四氮唑比色法、菌落形成、EdU掺入、流式细胞术、细胞伤口愈合和透孔试验评估了NF-κB p65对细胞增殖、迁移、侵袭和上皮-间质转化的生物学效应:结果:NF-κB p65 高表达,而 p-p65 Ser536 在中晚期 HCC 组织中表达不高。体内实验表明,RelA/p65 Ser536 的磷酸化模拟突变体(p65/S536D)能阻止肿瘤的进展和转移。体外实验表明,p65/S536D 可抑制肿瘤的增殖、迁移和侵袭。从机制上讲,RelA/p65 Ser536磷酸化抑制了NF-κB p65的核转位,并减少了NF-κB p65与BCL2、SNAIL和MMP9启动子的结合:结论:RelA/p65 Ser536磷酸化不利于NF-κB p65进入细胞核,并通过减少BCL2、SNAIL和MMP9抑制HCC的进展和转移。RelA/p65 Ser536的磷酸化位点极有可能成为HCC中NF-κB靶向治疗的潜在靶点。
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
Gastroenterology Report
Gastroenterology Report Medicine-Gastroenterology
CiteScore
4.60
自引率
2.80%
发文量
63
审稿时长
8 weeks
期刊介绍: Gastroenterology Report is an international fully open access (OA) online only journal, covering all areas related to gastrointestinal sciences, including studies of the alimentary tract, liver, biliary, pancreas, enteral nutrition and related fields. The journal aims to publish high quality research articles on both basic and clinical gastroenterology, authoritative reviews that bring together new advances in the field, as well as commentaries and highlight pieces that provide expert analysis of topical issues.
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