Bone morphogenetic protein 4 induces differentiation of colorectal cancer stem cells and increases their response to chemotherapy in mice

Gastroenterology. 2011 Jan;140(1):297-309. doi: 10.1053/j.gastro.2010.10.005. Epub 2010 Oct 14.

Abstract

Background & aims: The limited clinical response observed in many patients with colorectal cancer may be related to the presence of chemoresistant colorectal cancer stem cells (CRC-SCs). Bone morphogenetic protein 4 (BMP4) promotes the differentiation of normal colonic stem cells. We investigated whether BMP4 might be used to induce differentiation of CRC-SCs and for therapeutic purposes.

Methods: CRC-SCs were isolated from 25 tumor samples based on expression of CD133 or using a selection culture medium. BMP4 expression and activity on CRC-SCs were evaluated in vitro; progeny of the stem cells were evaluated by immunofluorescence, immunoblot, and flow cytometry analyses. The potential therapeutic effect of BMP4 was assessed in immunocompromised mice after injection of CRC-SCs that responded to chemotherapy (n = 4) or that did not (n = 2).

Results: CRC-SCs did not express BMP4 whereas differentiated cells did. Recombinant BMP4 promoted differentiation and apoptosis of CRC-SCs in 12 of 15 independent experiments; this effect did not depend on Small Mothers against decapentaplegic (Smad)4 expression level or microsatellite stability. BMP4 activated the canonical and noncanonical BMP signaling pathways, including phosphoInositide 3-kinase (PI3K) and PKB (protein kinase B)/AKT. Mutations in PI3K or loss of Phosphatase and Tensin homolog (PTEN) in Smad4-defective tumors made CRC-SCs unresponsive to BMP4. Administration of BMP4 to immunocompromised mice with tumors that arose from CRC-SCs increased the antitumor effects of 5-fluorouracil and oxaliplatin.

Conclusions: BMP4 promotes terminal differentiation, apoptosis, and chemosensitization of CRC-SCs in tumors that do not have simultaneous mutations in Smad4 and constitutive activation of PI3K. BMP4 might be developed as a therapeutic agent against cancer stem cells in advanced colorectal tumors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • AC133 Antigen
  • Adenomatous Polyposis Coli / genetics
  • Adenomatous Polyposis Coli / pathology
  • Aged
  • Aged, 80 and over
  • Animals
  • Antigens, CD / metabolism
  • Antineoplastic Agents / pharmacology
  • Antineoplastic Agents / therapeutic use*
  • Apoptosis / drug effects
  • Bone Morphogenetic Protein 4 / pharmacology
  • Bone Morphogenetic Protein 4 / therapeutic use*
  • Cell Differentiation
  • Cells, Cultured
  • Colorectal Neoplasms / drug therapy*
  • Colorectal Neoplasms / pathology
  • Female
  • Fluorouracil / therapeutic use*
  • Glycoproteins / metabolism
  • Humans
  • Male
  • Mice
  • Microsatellite Instability
  • Middle Aged
  • Mutation
  • Neoplastic Stem Cells / drug effects*
  • Organoplatinum Compounds / therapeutic use*
  • Oxaliplatin
  • PTEN Phosphohydrolase / genetics
  • PTEN Phosphohydrolase / metabolism
  • Peptides / metabolism
  • Phosphatidylinositol 3-Kinase / genetics
  • Phosphatidylinositol 3-Kinase / metabolism
  • Proto-Oncogene Proteins c-akt / metabolism
  • Smad4 Protein / metabolism

Substances

  • AC133 Antigen
  • Antigens, CD
  • Antineoplastic Agents
  • Bone Morphogenetic Protein 4
  • Glycoproteins
  • Organoplatinum Compounds
  • PROM1 protein, human
  • Peptides
  • Prom1 protein, mouse
  • SMAD4 protein, human
  • Smad4 Protein
  • Smad4 protein, mouse
  • Oxaliplatin
  • Phosphatidylinositol 3-Kinase
  • Proto-Oncogene Proteins c-akt
  • PTEN Phosphohydrolase
  • PTEN protein, human
  • Pten protein, mouse
  • Fluorouracil