Effect of DExH-box helicase 9 O-GlcNAc modification on the proliferation of HBV-associated hepatoma cells
10.13406/j.cnki.cyxb.003824
- VernacularTitle:DExH-box解旋酶9 O-GlcNAc修饰对HBV相关肝癌细胞增殖能力的影响
- Author:
Huimin LUO
1
;
Yubo PI
;
Yanmeng CHEN
;
Kai WANG
;
Ni TANG
;
Ailong HUANG
Author Information
1. 重庆医科大学感染性疾病分子生物学教育部重点实验室,重庆 400016
- Keywords:
DExH-box helicase 9;
O-linked N-acetylglucosamine;
hepatocellular carcinoma;
cell proliferation
- From:
Journal of Chongqing Medical University
2025;50(6):799-807
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of DExH-box helicase 9(DHX9)O-linked N-acetylglucosamine(O-GlcNAc)modifi-cation(O-GlcNAcylation)on the proliferation of hepatitis B virus(HBV)-associated hepatoma cells.Methods:The pAdTrack-TO4-DHX9-3Flag recombinant adenovirus plasmid was constructed by molecular cloning and transfected into HEK293 cells for packaging and amplification of the recombinant adenovirus AdDHX9.The interaction between DHX9 and O-GlcNAc transferase(OGT)was con-firmed using co-immunoprecipitation.The co-localization between DHX9 and OGT was measured by immunofluorescence.The level of DHX9 O-GlcNAcylation was determined using succinylated wheat germ agglutinin(sWGA)and glycosylated immunoprecipitation(IP).The effect of DHX9 O-GlcNAcylation on the proliferation of HBV-associated hepatoma cells was assessed using the colony-forming assay and cell growth curves.Results:The recombinant adenovirus AdDHX9 was successfully obtained,and DHX9 expression was confirmed by Western blot.DHX9 interacted with OGT,and the two proteins were co-localized on the nucleus.The sWGA and gly-cosylated IP experiments showed that DHX9 underwent O-GlcNAcylation,which was further enhanced by HBV infection.The colony-forming assay demonstrated that the number of cell clones was increased in the AdDHX9 group(386.667±15.630)compared with the AdGFP control group(142.667±7.572,P<0.001).Moreover,cell growth curves demonstrated that the overall cell growth rate was en-hanced in the AdDHX9 group(22.860±0.770)compared with the AdGFP control group(13.670±0.517,P<0.001).Conclusion:HBV infection promotes DHX9 O-GlcNAcylation,which enhances the proliferation of HBV-associated hepatoma cells.
