BRAFV600E-dependent Mcl-1 Stabilization Leads to Everolimus Resistance in Colon Cancer Cells

Oncotarget. 2016 Jul 26;7(30):47699-47710. doi: 10.18632/oncotarget.10277.

Abstract

mTOR activation is commonly caused by oncogenic mutations in RAS/RAF/MAPK and PI3K/AKT pathways, and promotes cancer progression and therapeutic resistance. However, mTOR inhibitors show limited single agent efficacy in patients. mTOR inhibitors suppress tumor cell growth and angiogenesis, and have recently been shown to induce death receptor/FADD-dependent apoptosis in colon cancers. Using a panel of BRAF V600E and WT colorectal cancer cell lines and in vitro selected resistant culture, and xenograft models, we demonstrate here that BRAFV600E confers resistance to mTOR inhibitors. Everolimus treatment disrupts the S6K1-IRS-2/PI3K negative feedback loop, leading to BRAF V600E-dependent activation of ERK and Mcl-1 stabilization in colon cancer cells, which in turn blocks the crosstalk from the death receptor to mitochondria. Co-treatment with inhibitors to Mcl-1, PI3K, RAF or MEK restores mTOR inhibitor-induced apoptosis by antagonizing Mcl-1 or abrogating ERK activation in BRAFV600E cells. Our findings provide a rationale for genotype-guided patient stratification and potential drug combinations to prevent or mitigate undesired activation of survival pathways induced by mTOR inhibitors.

Keywords: BRAF V600E; ERK; Mcl-1; everolimus; mTOR.

MeSH terms

  • Animals
  • Antineoplastic Agents / pharmacology
  • Cell Line, Tumor
  • Colorectal Neoplasms / drug therapy*
  • Colorectal Neoplasms / genetics
  • Colorectal Neoplasms / metabolism
  • Drug Resistance, Neoplasm
  • Everolimus / pharmacology*
  • Female
  • Humans
  • Mice
  • Mice, Nude
  • Myeloid Cell Leukemia Sequence 1 Protein / genetics
  • Myeloid Cell Leukemia Sequence 1 Protein / metabolism*
  • Protein Stability
  • Proto-Oncogene Proteins B-raf / genetics
  • Proto-Oncogene Proteins B-raf / metabolism*
  • Signal Transduction
  • TOR Serine-Threonine Kinases / antagonists & inhibitors
  • Transfection
  • Xenograft Model Antitumor Assays

Substances

  • Antineoplastic Agents
  • MCL1 protein, human
  • Myeloid Cell Leukemia Sequence 1 Protein
  • Everolimus
  • MTOR protein, human
  • TOR Serine-Threonine Kinases
  • BRAF protein, human
  • Proto-Oncogene Proteins B-raf