Sublethal doxorubicin promotes migration and invasion of breast cancer cells: role of Src Family non-receptor tyrosine kinases

Breast Cancer Research

Ronald S. Chavez¹  Samia Mohammed²  Yusuf A. Hannun³  Achraf A. Shamseddine⁴  Benjamin Newcomb⁴  Tyler D. Panzner⁵  Chioma M. Okeoma⁵  Daniel Canals⁶  Christopher J. Clarke⁶  Allen H. Lee⁷ 

[1] Department of Biochemistry and Cell Biology, Stony Brook University, 11794- 8430, Stony Brook, NY, USA;Department of Biochemistry and Cell Biology, Stony Brook University, 11794- 8430, Stony Brook, NY, USA;Stony Brook University Cancer Center, MART Level 9, Stony Brook University, 11794-8430, Stony Brook, NY, USA;Department of Medicine, Stony Brook University, Health Science Center, Hospital Pavilion Level 5, 11794-8430, Stony Brook, NY, USA;Department of Biochemistry and Cell Biology, Stony Brook University, 11794- 8430, Stony Brook, NY, USA;Stony Brook University Cancer Center, MART Level 9, Stony Brook University, 11794-8430, Stony Brook, NY, USA;Department of Medicine, Stony Brook University, Health Science Center, Hospital Pavilion Level 5, 11794-8430, Stony Brook, NY, USA;Department of Pharmacological Sciences, Stony Brook University, 11794-8430, Stony Brook, NY, USA;The Northport Veterans Affairs Hospital, 11768, Northport, NY, USA;Department of Medicine, Stony Brook University, Health Science Center, Hospital Pavilion Level 5, 11794-8430, Stony Brook, NY, USA;Department of Pharmacological Sciences, Stony Brook University, 11794-8430, Stony Brook, NY, USA;Stony Brook University Cancer Center, MART Level 9, Stony Brook University, 11794-8430, Stony Brook, NY, USA;Department of Medicine, Stony Brook University, Health Science Center, Hospital Pavilion Level 5, 11794-8430, Stony Brook, NY, USA;Stony Brook University Cancer Center, MART Level 9, Stony Brook University, 11794-8430, Stony Brook, NY, USA;Department of Medicine, Stony Brook University, Health Science Center, Hospital Pavilion Level 5, 11794-8430, Stony Brook, NY, USA;Department of Pharmacological Sciences, Stony Brook University, 11794-8430, Stony Brook, NY, USA;

关键词: Src Family Kinases;    Fyn;    Yes;    Doxorubicin;    p53;    ATR;    Dasatinib;    Breast cancer;   

DOI  :  10.1186/s13058-021-01452-5

来源: Springer

【 摘 要 】

BackgroundDoxorubicin (Dox) is a widely used chemotherapy, but its effectiveness is limited by dose-dependent side effects. Although lower Dox doses reduce this risk, studies have reported higher recurrence of local disease with no improvement in survival rate in patients receiving low doses of Dox. To effectively mitigate this, a better understanding of the adverse effects of suboptimal Dox doses is needed.MethodsEffects of sublethal dose of Dox on phenotypic changes were assessed with light and confocal microscopy. Migratory and invasive behavior were assessed by wound healing and transwell migration assays. MTT and LDH release assays were used to analyze cell growth and cytotoxicity. Flow cytometry was employed to detect cell surface markers of cancer stem cell population. Expression and activity of matrix metalloproteinases were probed with qRT-PCR and zymogen assay. To identify pathways affected by sublethal dose of Dox, exploratory RNAseq was performed and results were verified by qRT-PCR in multiple cell lines (MCF7, ZR75-1 and U-2OS). Regulation of Src Family kinases (SFK) by key players in DNA damage response was assessed by siRNA knockdown along with western blot and qRT-PCR. Dasatinib and siRNA for Fyn and Yes was employed to inhibit SFKs and verify their role in increased migration and invasion in MCF7 cells treated with sublethal doses of Dox.ResultsThe results show that sublethal Dox treatment leads to increased migration and invasion in otherwise non-invasive MCF7 breast cancer cells. Mechanistically, these effects were independent of the epithelial mesenchymal transition, were not due to increased cancer stem cell population, and were not observed with other chemotherapies. Instead, sublethal Dox induces expression of multiple SFK—including Fyn, Yes, and Src—partly in a p53 and ATR-dependent manner. These effects were validated in multiple cell lines. Functionally, inhibiting SFKs with Dasatinib and specific downregulation of Fyn suppressed Dox-induced migration and invasion of MCF7 cells.ConclusionsOverall, this study demonstrates that sublethal doses of Dox activate a pro-invasive, pro-migration program in cancer cells. Furthermore, by identifying SFKs as key mediators of these effects, our results define a potential therapeutic strategy to mitigate local invasion through co-treatment with Dasatinib.

【 授权许可】

CC BY