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Prevention of cisplatin-induced ototoxicity by the inhibition of gap junctional intercellular communication in auditory cells

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Abstract

Cis-diamminedichloroplatinum (cisplatin) is an effective chemotherapeutic drug for cancer therapy. However, most patients treated with cisplatin are at a high risk of ototoxicity, which causes severe hearing loss. Inspired by the “Good Samaritan effect” or “bystander effect” from gap junction coupling, we investigated the role of gap junctions in cisplatin-induced ototoxicity as a potential therapeutic method. We showed that connexin 43 (Cx43) was highly expressed in House Ear Institute-Organ of Corti 1 (HEI-OC1) cells, mediating cell–cell communication. The viability of HEI-OC1 cells was greatly decreased by cisplatin treatment, and cisplatin-treated HEI-OC1 cells showed lower Cx43 expression compared to that of untreated HEI-OC1 cells. In particular, high accumulation of Cx43 was observed around the nucleus of cisplatin-treated cells, whereas scattered punctuate expression of Cx43 was observed in the cytoplasm and membrane in normal cells, suggesting that cisplatin may interrupt the normal gap junction communication by inhibiting the trafficking of Cx43 to cell membranes in HEI-OC1 cells. Interestingly, we found that the inhibition of gap junction activity reduced cisplatin-induced apoptosis of auditory hair cells. Cx43 siRNA- or 18α-GA-treated HEI-OC1 cells showed higher cell viability compared to control HEI-OC1 cells during cisplatin treatment; this was also supported by fluorescence recovery after photobleaching studies. Inhibition of gap junction activity reduced recovery of calcein acetoxymethyl ester fluorescence compared to control cells. Additionally, analysis of the mechanisms involved demonstrated that highly activate extracellular signal-regulated kinase and protein kinase B, combined with inhibition of gap junctions may promote cell viability during cisplatin treatment.

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Abbreviations

18α-GA:

18 alpha-glycyrrhetinic acid

Akt:

Protein kinase B

ATP:

Adenosine triphosphate

cAMP:

Cyclic adenosine monophosphate

cGMP:

Cyclic guanosine monophosphate

Cisplatin:

Cis-diamminedichloroplatinum

Cx:

Connexin

ERK:

Extracellular signal-regulated kinase

FRAP:

Fluorescence recovery after photobleaching

HEI-OC1:

House Ear Institute-Organ of Corti 1

InsP3:

Inositol trisphosphate

MAPK:

Mitogen-activated protein kinases

NOX3:

NADPH oxidase 3

PKC:

Protein kinase C

ROS:

Reactive oxygen species

SLDT:

Scrape load dye transfer

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Acknowledgments

This work was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education Science and Technology (2010-0010678).

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Authors and Affiliations

  1. Department of Otolaryngology, Ajou University School of Medicine, San 5 Woncheon-dong, Yeongtong-gu, Suwon, 443-721, Republic of Korea

    Yeon Ju Kim, Chunjie Tian, Hye Jin Lim, Young Sun Kim & Yun-Hoon Choung

  2. Department of Biosystems & Biomaterials Science and Engineering, Seoul National University, Seoul, 151-742, Republic of Korea

    Jangho Kim & Jong Hoon Chung

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  1. Yeon Ju Kim
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Kim, Y.J., Kim, J., Tian, C. et al. Prevention of cisplatin-induced ototoxicity by the inhibition of gap junctional intercellular communication in auditory cells. Cell. Mol. Life Sci. 71, 3859–3871 (2014). https://doi.org/10.1007/s00018-014-1594-3

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