Lipocalin2 Protects Human Embryonic Kidney Cells against Cisplatin–Induced Genotoxicity

Document Type: Research article

Authors

1 Student Research Center, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences and Health Services, Isfahan, Iran.

2 Department of Toxicology and Pharmacology, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences and Health Services, Isfahan, Iran.

3 Medical Biotechnology Research Center, Paramedicine Faculty, Guilan University of Medical Sciences, Rasht, Iran.

4 Department of Pharmaceutical Biotechnology, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences and Health Services, Isfahan, Iran.

Abstract

Cisplatin is one of the most useful chemotherapeutics which performs its cytotoxic effect
via accumulation of platinum resulting in oxidative stress, and destruction of cell DNA. This
could probably cause secondary cancers in healthy tissues. Lipocalin2 (Lcn2) is a protein which
its expression is increased in oxidative stresses. Therefore, the present study was performed
to evaluate the protective effects of Lcn2 up-regulation on cisplatin genotoxicity. In order to
up-regulate Lcn2 expression, HEK293 cells were transfected with pcDNA3.1-Lcn2 vector.
Afterwards, stable cells consistently expressing Lcn2 were selected via screening with G418
antibiotic. Next, overexpression of Lcn2 was evaluated by RT-PCR and ELISA, comparing to
the control non-transfected cells. Then, in order to evaluate the cytoprotective effects of Lcn2
overexpression, transfected and non-transfected cells were subjected to cisplatin treatment
followed by MTT and alkaline Comet assays. RT-PCR and ELISA assays confirmed upregulation
of Lcn2 by the stable cells. MTT assay of the Lcn2 over-expressing cells showed
higher IC50 values comparing to the non-transfected cells. Furthermore, the Comet assay
confirmed Lcn2 protective effects on the cisplatin (1 μg/mL) induced genotoxicity. In the
present study, for the first time, we showed the protective effect of Lcn2 on cisplatin induced
genotoxicity. Therefore, one of the probable mechanisms of Lcn2 cytoprotctive effects under
oxidative stress conditions could be due to the prevention of genotoxicity. However, further
evaluations in this regard must be considered.

Keywords

Main Subjects