Abstract
Cisplatin is used widely for treatment of a variety of cancer diseases. Recently, however, the use of cisplatin is restricted because of its adverse effects such as hepatotoxicity. There is no study with current proteomics technology to evaluate cisplatin-induced hepatotoxicity, even if some studies have reported on the hepatotoxicity. In this study, proteomic as well as genomic analyses have been used for identification of proteins and genes that respond to cisplatin treatment in rat primary hepatocytes. To investigate the hepatotoxic effects of cisplatin, rat primary hepatocytes were treated with an IC(20) concentration for 24 h. From proteomic analysis based on label-free quantitation strategy, cisplatin induced 76 up-regulated and 19 down-regulated proteins among 325 distinct proteins. In the mRNA level, genomic analysis revealed 72 up-regulated and 385 down-regulated genes in the cisplatin-treated group. Based on these two analyses, 19 pathways were commonly altered, whereas seven pathways were identified only by proteomic analysis, and 19 pathways were identified only by genomic analysis. Overall, this study explained the mechanism of cisplatin-induced hepatotoxicity with two points of view: well known pathways including drug metabolism, fatty acid metabolism, and glycolysis/TCA cycle and little known pathways including urea cycle and inflammation metabolism, for hepatotoxicity of other toxic agents. Up-regulated proteins detected by proteomic analysis in the cisplatin-treated group: FBP1 (fructose 1,6-bisphosphatase 1), FASN (fatty acid synthase), CAT (catalase), PRDX1 (peroxiredoxin-1), HSPD1 (60-kDa heat shock protein), MDH2 (malate dehydrogenase 2), and ARG1 (arginase 1), and also down-regulated proteins in the cisplatin-treated group: TPM1 (tropomyosin 1), TPM3 (tropomyosin 3), and CTSB (cathepsin B), were confirmed by Western blot analysis. In addition, up-regulated mRNAs detected by microarray analysis in the cisplatin-treated group: GSTA2, GSTT2, YC2, TXNRD1, CYP2E1, CYP2C13, CYP2D1, ALDH17, ARG1, ARG2, and IL-6, and also down-regulated mRNAs: CYP2C12, CYP26B1, TPM1, and TPM3, were confirmed by RT-PCR analysis. In case of PRDX1, FASN, and ARG1, they were further confirmed by immunofluorescence analysis. Through the integrated proteomic and genomic approaches, the present study provides the first pathway map related to cisplatin-induced hepatotoxicity, which may provide new insight into the mechanism of hepatotoxicity.
Highlights
IntroductionAs a result of the limitations in the current knowledge regarding the mechanisms of hepatotoxicity, drug-induced liver injury (DILI) is still a major clinical problem [12, 13]
From the ‡Department of Molecular Medicine, Cell and Matrix Biology Research Institute, School of Medicine, Kyungpook National University, Daegu 700-422, Republic of Korea, the §Department of Pharmacology, School of Medicine, Kyungpook National University, Daegu 700-422, Republic of Korea, ¶D & P Biotech, School of Medicine, Kyungpook National University, Daegu 700-422, Republic of Korea, the College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul 151-742, Republic of Korea, and the **Institute of Chemistry, Academia Sinica, Taipei 11529, Taiwan
Rat primary hepatocytes without cisplatin (0.1% Me2SO) as a control formed healthy hepatocytes, whereas cisplatintreated primary hepatocytes died in a dose-dependent manner (Fig. 1)
Summary
As a result of the limitations in the current knowledge regarding the mechanisms of hepatotoxicity, drug-induced liver injury (DILI) is still a major clinical problem [12, 13]. With the recent availability of techniques such as LC-MS/ MS-based shotgun proteomics, which significantly increase the number of proteins identified, the more proteins related to drug toxicity mechanisms have been discovered [13]. The aim of this study was to investigate the molecular events causing the hepatotoxicity of cisplatin and the underlying toxic mechanisms of cisplatin using rat primary hepatocytes. Seven proteins and 11 genes were confirmed by Western blot and RT-PCR analysis, respectively, and three proteins were further confirmed by immunofluorescence analysis This result would provide novel insight into the possible pathway of cisplatin-induced hepatotoxicity
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
