Generation and Characterization of an Isogenic Cell Line Model of Radioresistant Esophageal Adenocarcinoma

Methods Mol Biol. 2023:2645:139-152. doi: 10.1007/978-1-0716-3056-3_6.

Abstract

Radiation therapy is a cornerstone of cancer treatment worldwide. Unfortunately, in many cases, it does not control tumor growth, and many tumors display treatment resistance. The molecular pathways leading to treatment resistance in cancer have been subject to research for many years. Isogenic cell lines with divergent radiosensitivities are an extremely useful tool to study the molecular mechanisms that underpin radioresistance in cancer research, as they reduce the genetic variation that is present in patient samples and cell lines of different origin, thus allowing the elucidation of molecular determinants of radioresponse. Here, we describe the process of generating an in vitro isogenic model of radioresistant esophageal adenocarcinoma by chronic irradiation of esophageal adenocarcinoma cells with clinically relevant doses of X-ray radiation. We also characterize cell cycle, apoptosis, reactive oxygen species (ROS) production, DNA damage and repair in this model to investigate the underlying molecular mechanisms of radioresistance in esophageal adenocarcinoma.

Keywords: Apoptosis; Cancer; Cell cycle; Clonogenic assay; DNA repair; Isogenic cell line; Radiation; Radioresistance; Reactive oxygen species.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma* / genetics
  • Adenocarcinoma* / metabolism
  • Adenocarcinoma* / radiotherapy
  • Apoptosis / radiation effects
  • Cell Line, Tumor
  • Esophageal Neoplasms* / genetics
  • Esophageal Neoplasms* / pathology
  • Esophageal Neoplasms* / radiotherapy
  • Humans
  • Radiation Tolerance / genetics

Supplementary concepts

  • Adenocarcinoma Of Esophagus