The protective effect of estrogen on cisplatin‐induced cytotoxicity of HK‐2 cells

Abstract

Introduction and hypothesisCisplatin is a widely used chemotherapeutic agent and has significant renal toxicity. Estrogen is a major sex hormone in females, and previous studies have shown that estrogen has many physiological protective effects. HK‐2 cell line is an immortalized proximal tubule epithelial cell line from healthy men. In this study, we investigated the protective roles of estrogen on cisplatin‐induced nephrotoxicity by the in vitro HK‐2 cell model.MethodsHK‐2 Cells were pretreated with 17β‐estradiol (0~20 nM), estrogen receptor agonists, and estrogen receptor antagonists for 24 hours respectively. All groups of cells were followed treated with cisplatin (0–15 mg/ml) for 24 hours. The cell survival rate, proliferation, morphology were analyzed by ELISA. The apoptosis/necrosis were analyzed by Annexin‐V/PI staining using flow cytometry. Estrogen receptors and apoptosis related proteins expressions were confirmed by Western blotting.ResultsAll three types of estrogen receptors could be expressed by the HK‐2 cells. In cisplatin‐induced nephrotoxicity cell model, 17β‐estradiol pretreatment have higher cell survival rate than vehicle when dose larger than 10 nM. The Annexin‐V positive apoptotic cells and caspase‐3 expressions in 17β‐estradiol pretreated group is significant lower than vehicle control. The three types of estrogen agonists exert the protective effects in cisplatin‐induced cytotoxicity, and acted on the G protein‐coupled estrogen receptor (GPER) is the most effective. In addition, blocked GPER also significantly accentuated cytotoxicity of cisplatin treated HK‐2 cells.ConclusionsOur data suggested that estrogen pretreatment could protect HK‐2 cells from cisplatin induced cytotoxicity, and this protective effect occurs mainly through the GPER pathway. It may offer a novel therapeutic strategy for cisplatin‐induced nephrotoxicity. Further investigation into the molecular mechanisms and clinical applications are needed.Support or Funding InformationKMU‐TP105E37This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.