INCREASING OF DOPAMINERGIC PC12 CELL SURVIVAL AGAINST 6-OHDA-MEDIATED TOXICITY FOLLOWING OVER EXPRESSION OF DJ-1 FACTOR

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LASEMI S.; GARDANEH M.; PARVIN AKBARI P.

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Journal: JOURNAL OF CELL & TISSUE Year:2016 | Volume:7 | Issue:2 Page(s): 141-148

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Title

INCREASING OF DOPAMINERGIC PC12 CELL SURVIVAL AGAINST 6-OHDA-MEDIATED TOXICITY FOLLOWING OVER EXPRESSION OF DJ-1 FACTOR

Author(s)

LASEMI S. | GARDANEH M. | PARVIN AKBARI P. | Issue Writer Certificate

Keywords

PARKINSONQ1

DJ-1Q1

DOPAMINERGICQ1

NEUROPROTECTIONQ1

Abstract

Aim: The purpose of this study is to examine DJ-1effect on increasing survival of DOPAMINERGIC (DAergic) cells against PARKINSONian toxicity.Material and Methods: First recombinant lenti-viruses transporters were produced with both DJ-1 and reporter Jred genes and were used to cells infection. To this end, three lentiviruses vectors namely transporter, packaging and envelope were applied for co-transfection of HEK-293T cells as virusproducing cell line.24 and 48 hours transfected cell media were collected and concentrated till lentivirus stock generation. Following transduction of DAergic PC12 cells with this concentrated virus stock, the infected cells overexpressed DJ-1. After treatment of these transduced cells with the 6-OHDA toxin, their survival rate was measured in comparison with control.Results: Transfection HEK-293T cells steps and PC12 cells transduction with the virus stock were done successful, because reporter Jred gene expression was observed using fluorescence microscope in both steps. Then DJ-1 overexpression was proofed using RT-PCR method. Next experiments indicated that DJ-1 overexpression causes significant increase in PC12 cells survival against produced toxicity of 6-OHDA. PC12 cell survival percentage that had DJ-1-overexpressing was 30% more than control cells survival percentage and this increasing was statistically significant.Conclusion: increasing of DJ-1 expression in DAergic PC12 cells significantly increased their resistance and perpetuity against 6-OHDA neurotoxicity.

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