Full-length LGR5-positive cells have chemoresistant characteristics in colorectal cancer

Br J Cancer. 2016 May 24;114(11):1251-60. doi: 10.1038/bjc.2016.112. Epub 2016 May 3.

Abstract

Background: Leucine-rich repeat-containing G-protein-coupled receptor 5 (LGR5) is a target of Wnt signalling and considered both a cancer stem cell marker and intestinal stem cell marker. We found first some splice variants of LGR5 in human intestine and elucidated the functional feature of full-length LGR5 (LGR5FL).

Methods: Reverse transcript PCR using mRNA extracted from intestine revealed the existence of LGR5 splice variants. We designed an antibody that recognises only LGR5FL and assessed immunohistochemically the distribution of LGR5FL-positive cells and Ki-67-positive cells in clinical samples.

Results: Two LGR5 splice variants were expressed in the human intestine crypt cells; one lacked exon 5 and the other lacked exons 5-8. Only LGR5FL appeared during cell cycle arrest, whereas the transcript variants appeared when the cell cycle was proceeding. Immunohistochemistry and in situ hybridisation showed that LGR5FL-positive cells were negative for Ki-67. Comparing prechemotherapy and post-chemotherapy specimens, the population of LGR5FL-positive cells significantly increased with therapy (P<0.01).

Conclusions: The function of LGR5FL-positive cells had low cell proliferative ability compared with the cells, which expressed splice variants of LGR5 and remained after chemotherapy. Designing therapeutic strategies that target LGR5FL-positive cells seems to be important in colorectal cancer.

MeSH terms

  • Adenocarcinoma / drug therapy
  • Adenocarcinoma / metabolism*
  • Adenocarcinoma / surgery
  • Alternative Splicing
  • Antineoplastic Combined Chemotherapy Protocols / therapeutic use*
  • Cell Count
  • Cell Cycle / genetics
  • Cell Division
  • Cell Line, Tumor
  • Cetuximab / administration & dosage
  • Colon / metabolism
  • Colorectal Neoplasms / drug therapy
  • Colorectal Neoplasms / metabolism*
  • Colorectal Neoplasms / surgery
  • Drug Resistance, Neoplasm / physiology*
  • Exons
  • Fluorouracil / administration & dosage
  • Gene Expression Regulation, Neoplastic
  • Humans
  • Ileum / metabolism
  • Intestinal Mucosa / metabolism
  • Ki-67 Antigen / analysis
  • Neoadjuvant Therapy
  • Neoplasm Proteins / chemistry
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / physiology*
  • Organoplatinum Compounds / administration & dosage
  • Oxaliplatin
  • Protein Isoforms / genetics
  • Protein Isoforms / physiology
  • Receptors, G-Protein-Coupled / chemistry
  • Receptors, G-Protein-Coupled / genetics
  • Receptors, G-Protein-Coupled / physiology*

Substances

  • Ki-67 Antigen
  • LGR5 protein, human
  • Neoplasm Proteins
  • Organoplatinum Compounds
  • Protein Isoforms
  • Receptors, G-Protein-Coupled
  • Oxaliplatin
  • Cetuximab
  • Fluorouracil