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  • Title: [Changes of nucleolin expression and cellular localization during HUVEC apoptosis induced by hydrogen peroxide].
    Author: Wang HY, Liu XL, Jiang BM.
    Journal: Zhong Nan Da Xue Xue Bao Yi Xue Ban; 2008 Jun; 33(6):488-93. PubMed ID: 18599995.
    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.
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