Combining Cell Type-Restricted Adenoviral Targeting with Immunostaining and Flow Cytometry to Identify Cells-of-Origin of Lung Cancer

Methods Mol Biol. 2018:1725:15-29. doi: 10.1007/978-1-4939-7568-6_2.

Abstract

Lung cancers display considerable intertumoral heterogeneity, leading to the classification of distinct tumor subtypes. Our understanding of the genetic aberrations that underlie tumor subtypes has been greatly enhanced by recent genomic sequencing studies and state-of-the-art gene targeting technologies, highlighting evidence that distinct lung cancer subtypes may be derived from different "cells-of-origin". Here, we describe the intra-tracheal delivery of cell type-restricted Ad5-Cre viruses into the lungs of adult mice, combined with immunohistochemical and flow cytometry strategies for the detection of lung cancer-initiating cells in vivo.

Keywords: Cells-of-origin; Flow cytometry; Immunohistochemical staining; Intra-tracheal injection; Recombinant adenovirus; Tissue dissociation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma / diagnosis*
  • Adenocarcinoma / genetics
  • Adenoviridae / genetics*
  • Animals
  • Disease Models, Animal
  • Flow Cytometry / methods*
  • Genetic Therapy
  • Genetic Vectors / administration & dosage*
  • Humans
  • Immunohistochemistry / methods*
  • Integrases / genetics
  • Integrases / metabolism
  • Lung Neoplasms / diagnosis*
  • Lung Neoplasms / genetics
  • Mice
  • Mice, Knockout
  • Mutation
  • Proto-Oncogene Proteins p21(ras) / genetics
  • Transduction, Genetic
  • Tumor Cells, Cultured
  • Tumor Suppressor Protein p53 / genetics

Substances

  • KRAS protein, human
  • TP53 protein, human
  • Tumor Suppressor Protein p53
  • Cre recombinase
  • Integrases
  • Proto-Oncogene Proteins p21(ras)