Original Article

Citation: Cell Death and Disease (2017) 8, e2584; doi:10.1038/cddis.2016.473
Published online 2 February 2017

Colon cancer cell treatment with rose bengal generates a protective immune response via immunogenic cell death

Jianzhong Qin1, Nicholas Kunda1,2, Guilin Qiao1, Jed F Calata1,2, Krunal Pardiwala1,2, Bellur S Prabhakar3 and Ajay V Maker1,2,3

  1. 1Department of Surgery, Division of Surgical Oncology, University of Illinois at Chicago, Chicago, IL, USA
  2. 2Creticos Cancer Center, Advocate Illinois Masonic Medical Center, Chicago, IL, USA
  3. 3Department of Microbiology and Immunology, University of Illinois at Chicago, Chicago, IL, USA

Correspondence: AV Maker, Department of of Surgery, Division of Surgical Oncology, Department of Microbiology and Immunology, University of Illinois at Chicago, 835 South Wolcott Avenue, MC790, Chicago, IL 60612, USA. E-mail: amaker@uic.edu

Received 2 November 2016; Revised 13 December 2016; Accepted 14 December 2016

Edited by H-U Simon



Immunotherapeutic approaches to manage patients with advanced gastrointestinal malignancies are desired; however, mechanisms to incite tumor-specific immune responses remain to be elucidated. Rose bengal (RB) is toxic at low concentrations to malignant cells and may induce damage-associated molecular patterns; therefore, we investigated its potential as an immunomodulator in colon cancer. Murine and human colon cancer lines were treated with RB (10% in saline/PV-10) for cell cycle, cell death, and apoptosis assays. Damage-associated molecular patterns were assessed with western blot, ELISA, and flow cytometry. In an immunocompetent murine model of colon cancer, we demonstrate that tumors regress upon RB treatment, and that RB induces cell death in colon cancer cells through G2/M growth arrest and predominantly necrosis. RB-treated colon cancer cells expressed distinct hallmarks of immunogenic cell death (ICD), including enhanced expression of calreticulin and heat-shock protein 90 on the cell surface, a decrease in intracellular ATP, and the release of HMGB1. To confirm the ICD phenotype, we vaccinated immunocompetent animals with syngeneic colon cancer cells treated with RB. RB-treated tumors served as a vaccine against subsequent challenge with the same CT26 colon cancer tumor cells, and vaccination with in vitro RB-treated cells resulted in slower tumor growth following inoculation with colon cancer cells, but not with syngeneic non-CT26 cancer cells, suggesting a specific antitumor immune response. In conclusion, RB serves as an inducer of ICD that contributes to enhanced specific antitumor immunity in colorectal cancer.