In-depth Identification of Pathways Related to Cisplatin-induced Hepatotoxicity through an Integrative Method Based on an Informatics-assisted Label-free Protein Quantitation and Microarray Gene Expression Approach

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 downregulated 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. Molecular & Cellular

The Evidence of HeLa Cell Apoptosis Induced with Tetraethylammonium Using Proteomics and Various Analytical Methods

“…Quantitative proteomics analysis reveals that PDIA3, DJ-1, and GSTO had significantly changed expression in the HeLa/CDDP-resistant group (61). Notably, these proteins also significant changed under CDDP inducement in rat primary hepatocytes (62), and in our study, they are considered as potential TEA biomarkers. It might be concluded that TEA induced HeLa apoptosis not only by blocking KChs but also by following previous and credible the mechanism, such as CDDP, initiating a similar or the same apoptotic biological pathway.…”

“…In addition, DJ-1, PDIA3, and GSTO were selected as differential proteins in TEA-treated groups; they are also regulated proteins in CDDPresistant HeLa cells studies (61) as well as in a rat primary hepatocyte study of CDDP toxicity (62), and some important differential proteins are discovered in the p53-dependent apoptosis pathway as described above. These results provide new details on the new molecular pathway involved in apoptosis and give a broader context of apoptotic events in therapeutic strategies.…”

Hyperin protects against cisplatin-induced liver injury in mice

“…Cisplatin is becoming increasingly a favor to clinical doctors attributed to its unique clinical benefit. Unfortunately, cisplatin therapy brings about undesirable side effects, including renal dysfunction 4 , ototoxicity 5 , nausea and vomiting 6 , and hepatotoxicity 7 . Hepatotoxicity is not considered to be a dose limiting toxicity for cisplatin, but liver toxicity can occur when it is administered at high doses 8 , which has been an obstacle for the further usage of cisplatin.…”

Biomarkers for drug-induced liver injury

“…Several studies have already demonstrated the usefulness of using specific DNA or RNA sequences in the circulation, such as transcripts highly enriched in the liver, as markers to reflect DILI. 11,67,91 The concentrations of these molecules are too low to conduct a global survey using traditional nucleic acid profiling approaches like microarray. Besides microRNA, due to its small repertoire, no systematic survey of the DNA or RNA spectrum in the circulation has been done.…”