A novel chalcone derivative suppresses melanoma cell growth through targeting Fyn/Stat3 pathway

Ling Tang,Xu Zhang,Keke Li,Jing Long,Cong Peng,Xiang Chen

doi:10.1186/s12935-020-01336-2

Ling Tang, Xu Zhang + Show 4 more

Open Access

https://doi.org/10.1186/s12935-020-01336-2

Copy DOI

Abstract

BackgroundFyn has been documented to have oncogenic features in multiple tumors, which might be a potential therapeutic target, however, few studies on the function role of Fyn and its specific inhibitors in melanoma.MethodsWe investigated the impacts of Fyn and its inhibitor Lj-1-60 on melanoma through bioinformatics analysis, western blot, cell viability, cell cycle and apoptosis and xenograft tumor model as well as immunohistochemical staining. Pull-down and in vitro kinase assay were used to demonstrate Lj-1-60 targeting Fyn. Transcriptome sequencing and RT-PCR were adopted to confirm the potential mechanisms of Lj-1-60 in melanoma.ResultsOur findings showed that Fyn was overexpressed in melanoma cells and knocked down of Fyn suppressed the proliferation of melanoma cells. To identify the potential inhibitors of Fyn, our in-house library including total of 111,277 chemicals was conducted to vitro screening, among those compounds, 83 inhibitors were further detected to explore the effect on melanoma cells growth and discovered a novel chalcone derivative Lj-1-60 that exhibited low cellular toxicity and high anti-tumor efficacy. Lj-1-60 directly was associated with Fyn and inhibited the Fyn kinase activity with Stat3 as substrate. What’s more, Lj-1-60 suppressed the proliferation of melanoma in vitro and in vivo through inducing cell cycle arrest and apoptosis. Moreover, the activation of Stat3 had also been abrogated both in Lj-1-60 treated melanoma cells or Fyn knocked down cells.ConclusionOur study revealed a novel Fyn inhibitor that could significantly suppress melanoma growth, which is a promising potential inhibitor for melanoma treatment.

Highlights

Fyn has been documented to have oncogenic features in multiple tumors, which might be a potential therapeutic target, few studies on the function role of Fyn and its specific inhibitors in melanoma
Fyn is overexpressed in melanoma cells and knockdown of Fyn induces apoptosis in melanoma cells To verify the relationship between the expression of Fyn and melanoma, the relative expression of Fyn between melanoma tissues and normal tissues was evaluated based on GEO databases (GSE114445 and GSE29359)
Our findings showed that knockdown of Fyn significantly inhibited melanoma cells proliferation, compared with shMock group, the rate of proliferation in shFyn#1 group reduced by 38.9%, and in shFyn#4 group reduced by 68.7% at 72 h in melanoma cell Sk-Mel-5

Summary

Introduction

Fyn has been documented to have oncogenic features in multiple tumors, which might be a potential therapeutic target, few studies on the function role of Fyn and its specific inhibitors in melanoma. Cutaneous melanoma is a fatal skin cancer whose worldwide incidence has sharply increased in recent years. Tang et al Cancer Cell Int (2020) 20:256 remarkable improvement in overall survival and progression-free survival [4,5,6]. Treatment with immune checkpoint inhibitors including anti-CTLA4 and antiPD-1 have been confirmed bringing extensive benefit for metastatic melanoma patients [7, 8]. There are some obvious limitations in targeted therapy and immunotherapy. Patients receiving BRAFi monotherapy or combination therapy with BRAFi and MEKi within 11 to 15 months resulted in drug resistance [9]. The efficacy of immunotherapy is approximately 20–30% even in melanoma [7]; It is essential that novel inhibitors are needed for melanoma treatment

Methods

Results

Conclusion