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Protective effect of klotho protein against cisplatin ototoxicity in an auditory cell line

  • Se Jin Park (a1), So Hyun Park (a2), Ji Won Chang (a1), June Choi (a1), Hak Hyun Jung (a1) (a2) and Gi Jung Im (a1)...



Klotho protein is involved in insulin-signalling and ageing. Klotho mutation causes premature ageing and significantly shortens the lifespan. The anti-neoplastic drug cisplatin promotes ototoxicity at higher doses by inducing apoptosis. This study aimed to clarify the effect of klotho expression on cisplatin ototoxicity, using an auditory cell line.

Materials and methods:

Expressions of klotho messenger RNA and protein were analysed by reverse-transcription polymerase chain reaction and western blotting. Auditory cells (HEI-OC1 line) were pretreated with 2 nM klotho protein for 2 hours; 15 µM cisplatin was then applied. After 48 hours incubation, assessment of cell viability (via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide assay), apoptosis (via Hoechst 33258 staining) and reactive oxygen species was performed.


Klotho protein expression increased in cisplatin-treated auditory cells. Cells treated with both klotho protein and cisplatin showed a viability of 67.7 per cent, versus 59.4 per cent in cisplatin-treated cells. Klotho significantly attenuated the cisplatin-induced increase in reactive oxygen species, and increased the viability of cells with cisplatin-induced cytotoxicity.


Klotho protein is protective against cisplatin-induced auditory cell cytotoxicity; inhibition of reactive oxygen species may be the main mechanism.


Corresponding author

Address for correspondence: Dr Gi Jung Im, Department of Otolaryngology–Head and Neck Surgery, Korea University College of Medicine, 126-1 Anamdong 5-ga, Seongbuk-ku, Seoul, 136–705, South Korea Fax: (82) 2 925 5233 E-mail:


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1Kuro-o, M. Klotho and aging. Biochim Biophys Acta 2009;1790:1049–58
2Kuro-o, M, Matsumura, Y, Aizawa, H, Kawaguchi, H, Suga, T, Utsugi, T et al. Mutation of the mouse klotho gene leads to a syndrome resembling ageing. Nature 1997;390:4551
3Bartke, A. Long-lived klotho mice: new insights into the roles of IGF-1 and insulin in aging. Trends Endocrinol Metab 2006;17:33–5
4Manya, H, Akasaka-Manya, K, Endo, T. Klotho protein deficiency and aging. Geriatr Gerontol Int 2010;10(suppl 1):S80–7
5Kurosu, H, Yamamoto, M, Clark, JD, Pastor, JV, Nandi, A, Gurnani, P et al. Suppression of aging in mice by the hormone klotho. Science 2005;309:1829–33
6Kamemori, M, Ohyama, Y, Kurabayashi, M, Takahashi, K, Nagai, R, Furuya, N. Expression of klotho protein in the inner ear. Hear Res 2002;171:103–10
7Rybak, LP. Mechanisms of cisplatin ototoxicity and progress in otoprotection. Curr Opin Otolaryngol Head Neck Surg 2007;15:364–9
8Im, GJ, Chang, JW, Choi, J, Chae, SW, Ko, EJ, Jung, HH. Protective effect of Korean red ginseng extract on cisplatin ototoxicity in HEI-OC1 auditory cells. Phytother Res 2010;24:614–21
9Arking, DE, Krebsova, A, Macek M, Sr, Macek, M Jr, Arking, A, Mian, IS et al. Association of human aging with a functional variant of klotho. Proc Natl Acad Sci U S A 2002;99:856–61
10Fukino, K, Suzuki, T, Saito, Y, Shindo, T, Amaki, T, Kurabayashi, M et al. Regulation of angiogenesis by the aging suppressor gene klotho. Biochem Biophys Res Commun 2002;293:332–7
11Yamamoto, M, Clark, JD, Pastor, JV, Gurnani, P, Nandi, A, Kurosu, H et al. Regulation of oxidative stress by the anti-aging hormone klotho. J Biol Chem 2005;280:38029–34
12Kalinec, GM, Webster, P, Lim, DJ, Kalinec, F. A cochlear cell line as an in vitro system for drug ototoxicity screening. Audiol Neurootol 2003;8:177–89
13Rybak, LP, Whitworth, CA, Mukherjea, D, Ramkumar, V. Mechanisms of cisplatin-induced ototoxicity and prevention. Hear Res 2007;226:157–67
14Tatar, M, Bartke, A, Antebi, A. The endocrine regulation of aging by insulin-like signals. Science 2003;299:1346–51
15Brunet, A, Bonni, A, Zigmond, MJ, Lin, MZ, Juo, P, Hu, LS et al. Akt promotes cell survival by phosphorylating and inhibiting a Forkhead transcription factor. Cell 1999;96:857–68
16Wang, Y, Sun, Z. Klotho gene delivery prevents the progression of spontaneous hypertension and renal damage. Hypertension 2009;54:810–17
17Xiao, NM, Zhang, YM, Zheng, Q, Gu, J. Klotho is a serum factor related to human aging. Chin Med J (Engl) 2004;117:742–7



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