Changes of nucleolin expression and cellular localization during HUVEC apoptosis induced by hydrogen peroxide.
- Author:
Hai-Yun WANG
1
;
Xiao-Liu LIU
;
Bi-Mei JIANG
Author Information
1. Department of Pathophysiology, Xiangya School of Medicine, Central South University, Changsha 410078, China.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
drug effects;
Cell Nucleus;
metabolism;
Cells, Cultured;
Cytoplasm;
metabolism;
Endothelial Cells;
cytology;
metabolism;
Humans;
Hydrogen Peroxide;
pharmacology;
Phosphoproteins;
biosynthesis;
genetics;
RNA, Messenger;
biosynthesis;
genetics;
RNA-Binding Proteins;
biosynthesis;
genetics;
Umbilical Veins;
cytology
- From:
Journal of Central South University(Medical Sciences)
2008;33(6):488-493
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the expression and cellular localization of nucleolin C23 during human umbilical vein endothelial cell (HUVEC) apoptosis induced by hydrogen peroxide (H(2)O(2)).
METHODS:Apoptosis of HUVEC was induced by exposure to 0.5 mmol/L H(2)O(2) for different periods and detected by flow cytometry and activity of caspase-3. The mRNA and protein expression of nucleolin were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot, respectively. The intracellular distribution of nucleolin was observed by indirect immunofluorescence.
RESULTS:The percentage of apoptotic cells was increased significantly after treatment with H(2)O(2) for 12, 24 and 36 hours. The activity of caspase-3 reached the peak after treatment with H(2)O(2) for 4 h. RT-PCR showed that nucleolin C23 mRNA was decreased after 2, 4, and 8 hours treatment with H(2)O(2). Western blot showed that C23 protein level was decreased after 12 hours with an additional cleft band of 80 kD appeared after 8 hours. Density analysis showed that the 80 kD cleft band increased in a time-dependent manner. Immunofluorescence analysis demonstrated that H(2)O(2)-induced C23 redistribution from the nucleus to the cytoplasm.
CONCLUSION:H(2)O(2) could induce apoptosis accompanying with C23-cleavage and C23-translocation from the nucleus to the cytoplasm.
