Zinc supplementation is required for the cytotoxic and immunogenic effects of chemotherapy in chemoresistant p53-functionally deficient cells

Abstract

Optimal tumor eradication often results from the death of malignant cells, as induced by chemotherapeutic agents, coupled to the induction of antitumor immune responses. However, cancer cells frequently become resistant to the cytotoxic activity of chemotherapy. The aim of the present study was to evaluate whether zinc dichloride (ZnCl₂), which was known to re-establish the chemosensitivity of cancer cells by reactivating p53, promotes immunogenic instances of cell death. We found that ZnCl₂, in combination with chemotherapeutic agents such as cisplatin and adriamycin (ADR), favors the apoptotic demise of chemoresistant cells, while cisplatin and ADR alone fail to do so. The co-culture of immature dendritic cells (DCs) with cancer cells succumbing to the co-administration of chemotherapy and ZnCl₂ led to DC activation, as indicated by the upregulation of the activation markers CD83 and CD86. In part, such process depended on cell death, as it was limited (but not abrogated) by the pan-caspase inhibitor Z-VAD-fmk. Moreover, DC activation relied on the ZnCl₂-induced exposure of calreticulin (CRT) on the surface of cancer cells, correlating with the phosphorylation of eukaryotic translation initiation factor 2α (eIF2α), a marker of endoplasmic reticulum stress. The siRNA-mediated knockdown of CRT as well as the inhibition of CRT exposure with brefeldin A strongly impaired DC maturation, indicating CRT translocation as induced by that ZnCl₂ is a key event in this setting. Altogether, these results suggest that ZnCl₂, has the potential to enhance the therapeutic effects of antineoplastic agents not only by improving their cytotoxic activity but also by promoting CRT exposure.

Keywords: :

• apoptosis
• calreticulin
• chemoresistance
• chemotherapy
• combination therapy
• dendritic cell activation
• immunogenicity
• p53 reactivation
• tumor cells
• ZnCl₂

Citation: Garufi A, Di Renzo L, Granato M, Faggioni A, D’Orazi G, Cirone M. Zinc supplementation is required for the cytotoxic and immunogenic effects of chemotherapy in chemoresistant p53-functionally deficient cells. OncoImmunology 2013; 2:e26198; 10.4161/onci.26198

Disclosure of Potential Conflicts of Interest

No potential conflicts of interest were disclosed.

Acknowledgments

This study was supported by grants from the Italian Association for Cancer Research (AIRC, IG 11377 to GD and IG 10265 to AF). We thank Gianluca Bossi for critical discussion and Sandro Valia, Valeria Conte, Marina Peddis, and Donatella D’Eliseo for technical assistance.