DMF JBC Manuscript_Rebuttal_Final no Blue_Merged.pdf (766.07 kB)
0/0

Dimethyl fumarate inhibits the nuclear factor κB pathway in breast cancer cells by covalent modification of p65 protein

Download (766.07 kB)
journal contribution
posted on 09.03.2017 by I. Kastrati, M.I. Siklos, E.L. Calderon-Gierszal
In breast tumors, activation of the nuclear factor κB (NFκB) pathway promotes survival, migration, invasion, angiogenesis, stem cell-like properties, and resistance to therapy - all pheno-types of aggressive disease where therapy options remain limited. Adding an anti-inflammatory/anti-NFκB agent to breast cancer treatment would be beneficial, but no such drug is approved as either a monotherapy or adjuvant therapy. To address this need, we examined whether dimethyl fumarate (DMF), an anti-inflammatory drug already in clinical use for multiple sclerosis, can inhibit the NFκB pathway. We found that DMF effectively blocks NFκB activity in multiple breast cancer cell lines and abrogates NFκB-dependent mammosphere formation, indicating that DMF has anti-cancer stem cell properties. In addition, DMF inhibits cell proliferation and significantly impairs xenograft tumor growth. Mechanistically, DMF prevents p65 nuclear translocation and attenuates its DNA binding activity but has no effect on upstream proteins in the NFκB pathway. Dimethyl succinate, the inactive analog of DMF that lacks the electrophilic double bond of fumarate, is unable to inhibit NFκB activity. Also, the cell-permeable thiol N-acetyl L-cysteine, reverses DMF inhibition of the NFκB pathway, supporting the notion that the electrophile, DMF, acts via covalent modification. To determine whether DMF interacts directly with p65, we synthesized and used a novel chemical probe of DMF by incorporating an alkyne functionality and found that DMF covalently modifies p65, with cysteine 38 being essential for the activity of DMF. These results establish DMF as an NFKB inhibitor with anti-tumor activity that may add therapeutic value in the treatment of aggressive breast cancers.

Funding

National Institutes of Health (NIH), R01 CA130932 and R01 CA200669 to JF, R01 CA121107 to GRJT, and by a postdoctoral fellowship grant from Susan G. Komen for the Cure to IK (PDF12229484).

History

Publisher Statement

© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Publisher

American Society for Biochemistry and Molecular Biology

Language

en_US

issn

0021-9258

Issue date

01/02/2016

Exports

Categories

Exports