Regulation of Hippo/YAP signaling pathway ameliorates cochlear hair cell injury by regulating ferroptosis

Xiaorong Niu; Peng Han; Junsong Liu; Zichen Chen; Xiaoyan Ma; Ting Zhang; Baiya Li; Xudong Ma

doi:10.1016/j.tice.2023.102051

Regulation of Hippo/YAP signaling pathway ameliorates cochlear hair cell injury by regulating ferroptosis

Tissue Cell. 2023 Jun:82:102051. doi: 10.1016/j.tice.2023.102051. Epub 2023 Feb 26.

Authors

Xiaorong Niu¹, Peng Han¹, Junsong Liu¹, Zichen Chen², Xiaoyan Ma¹, Ting Zhang¹, Baiya Li¹, Xudong Ma³

Affiliations

¹ Department of Otorhinolaryngology Head and Neck Surgery, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, Shaanxi, China.
² Department of Otorhinolaryngology Head and Neck Surgery, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710004, Shaanxi, China.
³ Department of Neurosurgery, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, Shaanxi, China. Electronic address: Maxudong_1122@163.com.

PMID: 36889225
DOI: 10.1016/j.tice.2023.102051

Abstract

Cisplatin, which is effective for the treatment of solid tumors, also can induce cochlear hair cell damage. Therefore, this study was intended to explore how Hippo/YAP signaling pathway affects the cochlear hair cell injury by regulating ferroptosis. After cisplatin induction, or LAT1-IN-1 (YAP activator) and verteporfin (YAP inhibitor) treatment or transfection, the viability of HEI-OC1 cells was detected by cell counting kit-8 (CCK-8) assay. The iron level and the levels of oxidative stress markers (ROS, MDA and 4-HNE) were analyzed by iron assay kit, reactive oxygen species (ROS), malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE) assay kits, respectively. The expression of ferritin light chain (FTL) in HEI-OC1 cells was detected by immunofluorescence and protein expressions of yes associated protein (YAP,) phosphorylated YAP (p-YAP), transferrin receptor (TFRC), glutathione peroxidase 4 (GPX4), acyl-CoA synthetase long-chain family member 4 (ACSL4) and solute carrier family 7 member 11 (SLC7A11) in HEI-OC1 cells were detected by western blot. The transcription of FTL and TFRC by YAP1 was verified by dual-luciferase reporter assay. The transfection efficiency of small interfering RNA (si-RNA) specific to FTL (siRNA-FTL) and TFRC (siRNA-TFRC) was confirmed by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). As a result, cisplatin inhibited the viability of HEI-OC1 cells by increasing free Fe²⁺ level and decreasing FTL level. LAT1-IN-1 promoted the viability of cisplatin-induced HEI-OC1 cells by suppressing oxidative stress level, free Fe²⁺ level, ferroptosis and increasing FTL level, while the effect of verteporfin was the opposite. YAP1 transcriptionally regulated the expression of FTL and TFRC. Inhibition of FTL suppressed the viability of cisplatin-induced HEI-OC1 cells by increasing oxidative stress level, free Fe²⁺ level, ferroptosis and decreasing FTL level, while the effect of TFRC inhibition was the opposite. In conclusion, YAP1 ameliorated cochlear hair cell injury by upregulating FTL and TFRC to suppress ferroptosis.

Keywords: Cochlear hair cell; Ferritin light chain; Ferroptosis; Transferrin receptor; YAP.

MeSH terms

Apoptosis
Cisplatin* / pharmacology
Ferroptosis*
Hair Cells, Auditory
RNA, Small Interfering / metabolism
Reactive Oxygen Species / metabolism
Signal Transduction
Verteporfin / metabolism
Verteporfin / pharmacology

Substances

Cisplatin
Reactive Oxygen Species
Verteporfin
RNA, Small Interfering