Mutant BRAF upregulates MCL-1 to confer apoptosis resistance that is reversed by MCL-1 antagonism and cobimetinib in colorectal cancer

Hisato Kawakami, Shengbing Huang, Krishnendu Pal, Shamit K. Dutta, Debabrata Mukhopadhyay, Frank A Sinicrope

Research output: Contribution to journalArticle

14 Scopus citations

Abstract

Oncogenic BRAFV600E mutations activate MAPK signaling and are associated with treatment resistance and poor prognosis in patients with colorectal cancer. In BRAFV600E-mutant colorectal cancers, treatment failure may be related to BRAFV600E-mediated apoptosis resistance that occurs by an as yet undefined mechanism. We found that BRAFV600E can upregulate anti-Apoptotic MCL-1 in a gene dose-dependent manner using colorectal cancer cell lines isogenic for BRAF. BRAFV600E-induced MCL-1 upregulation was confirmed by ectopic BRAFV600E expression that activated MEK/ERK signaling to phosphorylate (MCL-1Thr163) and stabilize MCL-1.Upregulation ofMCL-1 wasmediated byMEK/ERK shown by the ability of ERK siRNA to suppress MCL-1. Stabilization of MCL-1 by phosphorylation was shown by a phosphorylationmimicking mutant and an unphosphorylated MCL-1 mutant that decreased or increasedMCL-1 protein turnover, respectively. MEK/ ERK inhibition by cobimetinib suppressed MCL-1 expression/ phosphorylation and induced proapoptotic BIMto a greater extent than did vemurafenib in BRAFV600E cell lines. MCL-1 knockdown versus control shRNA significantly enhanced cobimetinib-induced apoptosis in vitro and in HT29 colon cancer xenografts. The smallmolecule MCL-1 inhibitor, A-1210477, also enhanced cobimetinib-induced apoptosis in vitro that was due to disruption of the interaction ofMCL-1 with proapoptoticBAK and BIM. Knockdown of BIM attenuated BAX, but not BAK, activation by cobimetinib plus A-1210477. In summary, BRAFV600E-mediatedMEK/ERK activation can upregulate MCL-1 by phosphorylation/stabilization to confer apoptosis resistance that can be reversed by MCL-1 antagonismcombined with cobimetinib, suggesting a novel therapeutic strategy against BRAFV600E-mutant CRCs.

Original languageEnglish (US)
Pages (from-to)3015-3027
Number of pages13
JournalMolecular Cancer Therapeutics
Volume15
Issue number12
DOIs
StatePublished - Dec 1 2016

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ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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