Title: Expression of DAP-5 in HK2 cell apoptosis induced by gentamicin
Abstract: Objective To reproduce a model of gentamicin induced apoptosis of human kidney epithelial cell (HK2), to observe the expression of death-associated protein 5 (DAP5) in HK2 cells, and to explore the relationship between PI3K/Akt/ mTOR signal pathway and DAP5. Methods HK2 cells were cultured with 3.2mg/ml gentamicin. Cell apoptosis was determined by DNA agarose gel electrophoresis, the apoptotic rate was determined with flow cytometry. Protein expressions of DAP5, Akt and p-Akt were assessed with Western blotting. Rapamycin was used to block PI3K/Akt/mTOR signal pathway in HK2 cell apoptosis model, and the change in DAP5 expression was observed. Results HK2 apoptosis appeared 24h after gentamicin treatment, and the apoptotic rate was found to be increased with the prolongation of gentamicin treatment. The apoptotic rates were 5.9%, 23.0% and 49.9%, respectively, at 24h, 36h and 72h after gentamicin treatment (P<0.05). At the same time, lysis of DAP5 protein occurred to form DAP5/p86, and the expressions of DAP5/p86 and p-Akt protein were also upregulated gradually in the process (P<0.05). However, generation of DAP5/p86 was reduced when PI3K/Akt/mTOR signal pathway was blocked by rapamycin during this period (P<0.05). Conclusion Gentamicin may induce apoptosis of HK2 cell in time-dependent manner. During the process of apoptosis of HK2 cells, lysis of DAP5 may occur to form DAP5/p86, and PI3K/Akt/mTOR signal pathway may positively regulate the expression of DAP5.
DOI: 10.11855/j.issn.0577-7402.2015.11.10
Publication Year: 2015
Publication Date: 2015-01-11
Language: en
Type: article
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