Potent USP10/13 antagonist spautin-1 suppresses melanoma growth via ROS-mediated DNA damage and exhibits synergy with cisplatin.

Jia Guo,Jiangling Zhang,Min Qi,Cong Peng,Juan Su,Jing Liu,Long Liang,Xiang Chen,Nian Liu,Hong Liu,Shuang Zhao

doi:10.1111/jcmm.15093

Abstract

Malignant melanoma is one of the most invasive tumours. However, effective therapeutic strategies are limited, and overall survival rates remain low. By utilizing transcriptomic profiling, tissue array and molecular biology, we revealed that two key ubiquitin‐specific proteases (USPs), ubiquitin‐specific peptidase10 (USP10) and ubiquitin‐specific peptidase10 (USP13), were significantly elevated in melanoma at the mRNA and protein levels. Spautin‐1 has been reported as a USP10 and USP13 antagonist, and we demonstrated that spautin‐1 has potent anti‐tumour effects as reflected by MTS and the colony formation assays in various melanoma cell lines without cytotoxic effects in HaCaT and JB6 cell lines. Mechanistically, we identified apoptosis and ROS‐mediated DNA damage as critical mechanisms underlying the spautin‐1‐mediated anti‐tumour effect by utilizing transcriptomics, qRT‐PCR validation, flow cytometry, Western blotting and immunofluorescence staining. Importantly, by screening spautin‐1 with targeted or chemotherapeutic drugs, we showed that spautin‐1 exhibited synergy with cisplatin in the treatment of melanoma. Pre‐clinically, we demonstrated that spautin‐1 significantly attenuated tumour growth in a cell line‐derived xenograft mouse model, and its anti‐tumour effect was further enhanced by cotreatment with cisplatin. Taken together, our study revealed a novel molecular mechanism of spautin‐1 effecting in melanoma and identified a potential therapeutic strategy in treatment of melanoma patients.

Highlights

Malignant melanoma, a solid tumour produced by the malignant transformation of melanocytes from the skin and other organs, has an increasing incidence and a poor prognosis.[1]
Recent studies have shown that ubiquitin-specific peptidase10 (USP10) and ubiquitin-specific peptidase 13 (USP13) function as tumour promoters in human cancer by stabilizing multiple molecules.[24]
We discovered the high expression levels of USP10 and USP13 in malignant melanoma tissues and explored the function and underlying mechanisms of the anti-melanoma effects of the USP10/USP13 inhibitor spautin-1 through inducing the Reactive oxygen species (ROS)-mediated DNA damage

Summary

| INTRODUCTION

A solid tumour produced by the malignant transformation of melanocytes from the skin and other organs, has an increasing incidence and a poor prognosis.[1]. Chemotherapeutics or targeted therapeutics alter tumour growth, in most cases, the effect is not long-lasting resulting in drug resistance that causes treatment failure and impacts the survival of patients.[3,4] New immunotherapies have been recently approved to induce long-lasting responses in patients with metastatic cancers, even so, resistance and poor response rates still persist.[5] As melanoma eventually becomes resistant to these novel therapies, the development of more effective approaches is required. The advent of some new inhibitors revealed suppressive effect in melanoma.[8,9] Ubiquitin-specific peptidase[10] (USP10) and ubiquitin-specific peptidase 13 (USP13) both belong to USP family, the largest subfamily of DUB.[10] USP10 and USP13 are involved in a number of biological processes and impact the development of various tumours by stabilizing several proteins. Spautin-1 treatment could combine with cisplatin, a well-known chemotherapeutic drug, which was shown to enhance the anticancer effect of cisplatin both in vivo and in vitro

| MATERIALS AND METHODS

Findings

| DISCUSSION