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

作　　者：Wentao Zuo Haoyang Ma Jianghui Bi Tiaolan Li Yifeng Mo Shiyu Yu Jia Wang Beiqing Li Jinfeng Huang Yongwen Li Li Li 

机构地区：[1]College of Basic Medical,Guilin Medical University,Guilin,Guangxi,P.R.China 2College of Pharmacy,Guilin Medical University,Guilin,Guangxi,P.R.China

出　　处：《Gastroenterology Report》2024年第1期435-448,共14页胃肠病学报道(英文)

基　　金：funded by the College Students'Innovative Entrepreneurial Training Plan Program[no.202110601019].

摘　　要：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.

关 键 词：HCC NF-ΚB RelA/p65 Ser536 phosphorylation apoptosis METASTASIS 

分 类 号：R73[医药卫生—肿瘤]