Lipocalin2 Protects Human Embryonic Kidney Cells against Cisplatin–Induced Genotoxicity

Abstract

Cisplatin is one of the most useful chemotherapeutics which performs its cytotoxic effectvia accumulation of platinum resulting in oxidative stress, and destruction of cell DNA. Thiscould probably cause secondary cancers in healthy tissues. Lipocalin2 (Lcn2) is a protein whichits expression is increased in oxidative stresses. Therefore, the present study was performedto evaluate the protective effects of Lcn2 up-regulation on cisplatin genotoxicity. In order toup-regulate Lcn2 expression, HEK293 cells were transfected with pcDNA3.1-Lcn2 vector.Afterwards, stable cells consistently expressing Lcn2 were selected via screening with G418antibiotic. Next, overexpression of Lcn2 was evaluated by RT-PCR and ELISA, comparing tothe control non-transfected cells. Then, in order to evaluate the cytoprotective effects of Lcn2overexpression, transfected and non-transfected cells were subjected to cisplatin treatmentfollowed by MTT and alkaline Comet assays. RT-PCR and ELISA assays confirmed upregulationof Lcn2 by the stable cells. MTT assay of the Lcn2 over-expressing cells showedhigher IC50 values comparing to the non-transfected cells. Furthermore, the Comet assayconfirmed Lcn2 protective effects on the cisplatin (1 μg/mL) induced genotoxicity. In thepresent study, for the first time, we showed the protective effect of Lcn2 on cisplatin inducedgenotoxicity. Therefore, one of the probable mechanisms of Lcn2 cytoprotctive effects underoxidative stress conditions could be due to the prevention of genotoxicity. However, furtherevaluations in this regard must be considered.