[논문]Annexin A5 as a New Potential Biomarker for Cisplatin-Induced Toxicity in Human Kidney Epithelial Cells

Kwon, Yeo-Jung; Jung, Jin-Joo; Park, Na-Hee; Ye, Dong-Jin; Kim, Donghak; Moon, Aree; Chun, Young-Jin

doi:10.4062/biomolther.2013.026

Annexin A5 as a New Potential Biomarker for Cisplatin-Induced Toxicity in Human Kidney Epithelial Cells 원문보기

Biomolecules & therapeutics, v.21 no.3, 2013년, pp.190 - 195

Kwon, Yeo-Jung (College of Pharmacy, Chung-Ang University) , Jung, Jin-Joo (College of Pharmacy, Chung-Ang University) , Park, Na-Hee (College of Pharmacy, Chung-Ang University) , Ye, Dong-Jin (College of Pharmacy, Chung-Ang University) , Kim, Donghak (Department of Biological Sciences, Konkuk University) , Moon, Aree (College of Pharmacy, Duksung Women's University) , Chun, Young-Jin (College of Pharmacy, Chung-Ang University)

Abstract ▼ AI-Helper

Cisplatin is a member of platinum-containing anti-cancer drugs that causes cross-linking of DNA and ultimately cancer cell apoptosis. The therapeutic function of cisplatin on various types of cancers has been widely reported but the side effects have been discovered together and nephrotoxicity has been regarded as major side effect of cisplatin. To select candidates for new sensitive nephrotoxicity biomarker, we performed proteomic analysis using 2-DE/MALDI-TOF-MS followed by cisplatin treatment in human kidney cell line, HK-2 cells, and compared the results to the gene profile from microarray composed of genes changed in expression by cisplatin from formerly reported article. Annexin A5 has been selected to be the most potential candidate and it has been identified using Western blot, RT-PCR and cell viability assay whether annexin A5 is available to be a sensitive nephrotoxic biomarker. Treatment with cisplatin on HK-2 cells caused the increase of annexin A5 expression in protein and mRNA levels. Over-expression of annexin A5 blocked HK-2 cell proliferation, indicating correlation between annexin A5 and renal cell toxicity. Taken together, these results suggest the possibility of annexin A5 as a new biomarker for cisplatin-mediated nephrotoxicity.

주제어

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문제 정의

In this study, we explored a new biomarker for cisplatin-induced nephrotoxicity. To select proteins that show changes in expression followed by cisplatin treatment, proteomic analysis using 2DE/MALDI-TOF-MS was applied.

제안 방법

, 2008). In this paper, male rats were intravenously treated with cisplatin for 1, 3 and 5 days. Various types of genes reported to be associated with cisplatin-induced nephrotoxicity were examined and their level of change in expression were profi led.
In present study, we performed experiments to find new sensitive biomarker for nephrotoxicity. Screening for selection of candidates, proteomic analysis was performed. Because annexin A5 was selected as one of the most potential candidates, validation of sensitivity of annexin A5 for cisplatin-induced nephrotoxicity has also been implemented.

대상 데이터

Cisplatin was purchased from Sigma-Aldrich (St. Louis, MO, USA). DMEM medium, penicillin or streptomycin was obtained from Welgene (Daegu, Korea).
From the results of proteomic analysis, 160 spots showing significant increase of expression were selected as candidates for nephrotoxic biomarkers. To identify whether the corresponding results were observed in vivo, we compared proteins each spots represented to the previously reported DNA microarray results (Wang et al.
Immortalized human kidney epithelial cells (HK-2) was obtained from American Type Culture Collection (ATCC, Manassa, VA, USA) and cultured in DMEM medium supplemented with 10% (v/v) heat-inactivated FBS, 100 unit/ml penicillin, and 100 μg/ml streptomycin.

데이터처리

Using one-way analysis of variance, statistical analysis was performed followed by Dunnett's Multiple Comparison t-test using Graph-Pad Prism Software (GraphPad Software Inc., San Diego, CA) when appropriate.

후속연구

Due to the common property of annexin A4 and A5 that both annexins are more closely related to ion channel regulation than other annexins, the effects of cisplatin-induced annexin A4 and A5 on cell viability may be linked with their ability as regulator of ion channels. Further studies need to be conducted to identify the exact mechanism exploring the contribution of annexin A5 to renal cell apoptosis and whether annexin A5 could be used as general biomarker for wide range of renal failure.

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