Live Cell Imaging of Spatiotemporal Ca2+ Fluctuation Responses to Anticancer Drugs

Methods Mol Biol. 2022:2488:227-236. doi: 10.1007/978-1-0716-2277-3_15.

Abstract

Synthetic triterpenoids, 2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid (CDDO) and its derivatives are known to have potent anti-cancer and anti-inflammatory activities. The mechanisms of actions of CDDO and its derivatives as potential therapeutics remain elusive. Previous studies found that CDDO-Me triggers apoptosis by inducing extracellular Ca2+ influx followed by endoplasmic reticulum (ER)-derived vacuolation. Since Ca2+ activity in cells is dynamic and needs to be tracked in real time in living cells, we report a high-throughput and high-content imaging method to track CDDO-induced Ca2+ fluctuation in both ER and cytosol with MATLAB script for data analysis and visualization.

Keywords: Biosensor; CDDO; FRET; High-content analysis; High-throughput imaging.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Antineoplastic Agents* / pharmacology
  • Apoptosis
  • Oleanolic Acid* / analogs & derivatives
  • Oleanolic Acid* / pharmacology
  • Triterpenes*

Substances

  • Antineoplastic Agents
  • Triterpenes
  • Oleanolic Acid
  • bardoxolone