Assessment of quality and consistency of monoclonal antibodies for CB1 and CB2 in head and neck squamous cell carcinoma

Head Neck. 2019 Sep;41(9):3105-3113. doi: 10.1002/hed.25794. Epub 2019 Apr 29.

Abstract

Background: Marijuana has numerous roles as an agonist in the endocannabinoid signaling system (ESS). This study evaluated monoclonal antibodies across experimental techniques to establish a framework for studying ESS receptors, CB1 and CB2.

Methods: Tissue from five patients with head and neck cancer were used to generate cell lines and formalin-fixed paraffin-embedded (FFPE) sections, which were analyzed by western blot (WB), immunohistochemistry (IHC), and immunofluorescence (IF). Subgroup analysis was performed on FFPE sections from 8 marijuana users and 10 controls by IHC. Results were compared across methods for consistency.

Results: In all patients, WB and IF were CB1 positive, whereas IHC was negative. Select samples were CB2 positive by WB, but failed IF and IHC applications. In subgroup analysis, 1 of 8 users and 3 of 10 nonusers were CB1 positive.

Conclusions: Interpretation of CB1/CB2 antibody data should be performed cautiously and confirmation of findings across multiple experimental methods is recommended.

Keywords: CB1 receptor; CB2 receptor; endocannabinoid; head and neck; squamous cell carcinoma.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Antibodies, Monoclonal / metabolism*
  • Biopsy
  • Blotting, Western
  • Cannabis / immunology*
  • Case-Control Studies
  • Cell Line, Tumor
  • Female
  • Fluorescent Antibody Technique
  • Head and Neck Neoplasms / immunology*
  • Humans
  • Immunohistochemistry
  • Male
  • Middle Aged
  • Receptor, Cannabinoid, CB1 / metabolism*
  • Receptor, Cannabinoid, CB2 / metabolism*
  • Squamous Cell Carcinoma of Head and Neck / immunology*

Substances

  • Antibodies, Monoclonal
  • Receptor, Cannabinoid, CB1
  • Receptor, Cannabinoid, CB2