Up-regulated expression of neurotrophin-3 mediated by hypoxia response element attenuates apoptosis induced by hypoxia in PC12 cells

doi:10.16098/j.issn.0529-1356.2015.05.001

Abstract

Abstract:

Objective To investigate the controlled expression of neurotrophin-3 (NT-3) by HRE under hypoxic conditions and determine the protective effects of conditionally expressed NT-3 on hypoxia-induced apoptosis in PC12 cells. Methods Five copies of the HRE (5HRE) and NT-3 were employed to construct a therapeutic vector, and transferred into PC12 cells. Expression and secretion of NT-3 were detected by ELISA. Apoptosis of PC12 cells induced by hypoxia was assayed by TUNEL. Activation of p-38 and Caspase-3 was detected by Western blotting. Results The retroviral vectors were successfully constructed and transfected into PC12 cells to produce gene transferred cells, PC12-NT3-EGFP, PC12-5HRE-NT3-EGFP and PC12-5HRE-EGFP. Compared with normal conditions, in which NT-3 was expressed at low levels, the expression of NT-3 significantly increased under hypoxic conditions in PC12-5HRE-NT3-EGFP (n=3, P<0.05). The conditional adjustment of NT-3 expression by 5HRE significantly reduced apoptosis induced by hypoxia in PC12-5HRE-NT3-EGFP (n=3, P<0.05). In addition, the hypoxiainduced phosphorylation of both p38 and Caspase-3 activities was decreased in PC12-5HRE-NT3-EGFP under hypoxic conditions (n=3, P<0.05). Conclusion Up-regulated expression of NT-3 mediated by hypoxia response element in response to hypoxia in PC12 cells can protect PC12 cells against hypoxia-induced apoptosis.

Key words: Hypoxia response element, Neurotrophin-3, Hypoxia, Western blotting

ZHANG Jun-feng SHI Li-li ZHANG Li ZHAO Zhao-hua YANG Peng-bo ZHANG Jian-shui LIU Yong XU Xi*. Up-regulated expression of neurotrophin-3 mediated by hypoxia response element attenuates apoptosis induced by hypoxia in PC12 cells[J]. AAS, 2015, 46(5): 581-586.

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