AURKB inhibition induces rhabdomyosarcoma apoptosis and ferroptosis through NPM1/SP1/ACSL5 axis
Huimou Chen, Mengzhen Li, Yu Zhang, Mengjia Song, Yi Que, Juan Wang, Feifei Sun, Jia Zhu, Junting Huang, Juan Liu, Jiaqian Xu, Suying Lu, Yizhuo Zhang
Huimou Chen, Mengzhen Li, Yu Zhang, Mengjia Song, Yi Que, Juan Wang, Feifei Sun, Jia Zhu, Junting Huang, Juan Liu, Jiaqian Xu, Suying Lu, Yizhuo Zhang
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Research Article Oncology

AURKB inhibition induces rhabdomyosarcoma apoptosis and ferroptosis through NPM1/SP1/ACSL5 axis

Abstract

Rhabdomyosarcoma (RMS) is one of the most common solid tumors in children and adolescents. Patients with relapsed/refractory RMS have limited treatment options, highlighting the urgency for the identification of novel therapeutic targets for RMS. In the present study, aurora kinase B (AURKB) was found to be highly expressed in RMS and associated with unfavorable prognosis of patients. Functional experiments indicated that inhibition of AURKB significantly reduced RMS cell proliferation, induced apoptosis and ferroptosis, and suppressed RMS growth in vivo. The highly expressed AURKB in RMS contributes to the apoptosis and ferroptosis resistance of tumor cells through the nucleophosmin 1 (NPM1)/Sp1 transcription factor (SP1)/acyl-CoA synthetase long-chain family member 5 (ACSL5) axis. Furthermore, inhibition of AURKB exerted an anti-RMS effect together with vincristine both in vitro and in vivo, with tolerable toxicity. The above findings provide insights we believe are new into the tumorigenesis of RMS, especially with regard to apoptosis or ferroptosis resistance, indicating that AURKB may be a potential target for clinical intervention in patients with RMS.

Authors

Huimou Chen, Mengzhen Li, Yu Zhang, Mengjia Song, Yi Que, Juan Wang, Feifei Sun, Jia Zhu, Junting Huang, Juan Liu, Jiaqian Xu, Suying Lu, Yizhuo Zhang

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Figure 5

AURKB promoted apoptosis and ferroptosis resistance of RMS via phosphorylating Ser125 of NPM1.

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AURKB promoted apoptosis and ferroptosis resistance of RMS via phosphory...
(A) Silver staining after coimmunoprecipitation with AURKB antibody. The arrow denotes the distinct bands between 35 kDa and 40 kDa. (B) Results of protein mass spectrometry indicated the physical interaction of AURKB and NPM1. (C) Results of immunoprecipitation to verify the interaction of AURKB and NPM1. (D) Time-course analysis of WB identified expression changes of p-NPM1 at Ser125, Ser4, Thr95, Thr199, and the total NPM1 in RMS lines treated with AZD1152. (E) WB analysis identified expression changes of the p-NPM1 at Ser125, Ser4, Thr95, Thr199, and the total NPM1 in RMS lines after genetic knockdown of NPM1. (F) Colony formation assay after genetic knockdown of NPM1 with shRNA in RMS cell lines. (G) Colony formation assay evaluating proliferation ability after shRNA knockdown of NPM1 in RMS cells treated with or without apoptosis inhibitor (10 μM Z-VAD-FMK) or ferroptosis inhibitor (10 μM deferoxamine) for 48 hours. (H) Cell viability evaluated in RD and RH30 cell lines after knocking down NPM1 with shRNA in RMS cells treated with or without Z-VAD-FMK or deferoxamine treatment for 48 hours. (I) The apoptosis rate changes after NPM1 genetic knockdown with siRNA transfected for 72 hours in RMS cells. (J) Changes of the intracellular lipid ROS levels after NPM1 suppression with siRNA transfected for 72 hours. (K) Changes of the intracellular MDA levels after NPM1 suppression with siRNA transfected for 72 hours. (L) Changes of relative intracellular Fe2+ levels in RMS cells after NPM1 was silenced by siRNA transfection for 72 hours. (M) WB analysis revealed the changes of indicated apoptosis-related gene markers and the ferroptosis-related marker GPX4 after NPM1 inhibition with shRNA in RMS cells. **P < 0.01, ***P < 0.001, ****P < 0.0001, 2-way ANOVA (F–H) and t test (I–L).