Document Detail


Effect of 5-aza-2'-deoxycytidine on immune-associated proteins in exosomes from hepatoma.
MedLine Citation:
PMID:  20480522     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
AIM: To study the effect of 5-aza-2'-deoxycytidine (5-aza-CdR) on heat shock protein 70 (HSP70), human leucocyte antigen-I (HLA-I) and NY-ESO-1 proteins in exosomes produced by hepatoma cells, HepG2 and Hep3B. METHODS: Exosomes derived from HepG(2) and Hep3B cells treated with or without 5-aza-CdR were isolated and purified by ultrafiltration centrifugation and sucrose gradient ultracentrifugation. The number of exosomes was counted under electron microscope. Concentration of proteins in exosomes was measured by bicinchoninic acid protein assay. Expression of HSP70, HLA-I and NY-ESO-1 proteins in exosomes was detected by Western blotting and immunoelectron microscopy. mRNA expression of p53 gene was detected by reverse transcription polymerase chain reaction. RESULTS: The mRNA expression of p53 gene was increased in both hepatoma cell lines after treatment with 5-aza-CdR. The number of exosomes and the concentration of total proteins in exosomes were increased significantly after treatment with 5-aza-CdR (P < 0.05). After treatment with 5-aza-CdR, immunoelectron microscopy and Western blotting showed that the HSP70, HLA-I and NY-ESO-1 proteins were increased in exosomes produced by both hepatoma cell lines. CONCLUSION: 5-Aza-CdR, an inhibitor of DNA methyltransferase, can increase exosomes produced by hepatoma cells and immune-associated protein component of exosomes, which may be mediated by p53 gene up-regulation and 5-aza-CdR demethylation.
Authors:
Wen-Hua Xiao; Gao-Wa Sanren; Jian-Hua Zhu; Qiu-Wen Li; Huan-Rong Kang; Ru-Liang Wang; Lin-Ping Song; Ming Ye
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  World journal of gastroenterology : WJG     Volume:  16     ISSN:  1007-9327     ISO Abbreviation:  World J. Gastroenterol.     Publication Date:  2010 May 
Date Detail:
Created Date:  2010-05-18     Completed Date:  2010-08-26     Revised Date:  2010-09-30    
Medline Journal Info:
Nlm Unique ID:  100883448     Medline TA:  World J Gastroenterol     Country:  China    
Other Details:
Languages:  eng     Pagination:  2371-7     Citation Subset:  IM    
Affiliation:
Department of Oncology, First Affiliated Hospital, Chinese PLA General Hospital, Beijing 100048, China. w_hxiao@hotmail.com
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MeSH Terms
Descriptor/Qualifier:
Antigens, Neoplasm / metabolism
Azacitidine / analogs & derivatives*,  pharmacology
Blotting, Western
Carcinoma, Hepatocellular / enzymology,  genetics,  immunology*
Centrifugation, Density Gradient
DNA Modification Methylases / antagonists & inhibitors*,  metabolism
Enzyme Inhibitors / pharmacology*
Exosomes / drug effects*,  enzymology,  immunology
HLA Antigens / metabolism
HSP70 Heat-Shock Proteins / metabolism
Hep G2 Cells
Humans
Liver Neoplasms / enzymology,  genetics,  immunology*
Membrane Proteins / metabolism
Microscopy, Immunoelectron
RNA, Messenger / metabolism
Reverse Transcriptase Polymerase Chain Reaction
Tumor Suppressor Protein p53 / genetics
Up-Regulation
Chemical
Reg. No./Substance:
0/Antigens, Neoplasm; 0/CTAG1B protein, human; 0/Enzyme Inhibitors; 0/HLA Antigens; 0/HSP70 Heat-Shock Proteins; 0/Membrane Proteins; 0/RNA, Messenger; 0/TP53 protein, human; 0/Tumor Suppressor Protein p53; 2353-33-5/decitabine; 320-67-2/Azacitidine; EC 2.1.1.-/DNA Modification Methylases
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