The growing incidence of skin cancer (SC) is a global public health concern. Preventive approaches mostly embrace the sunscreen use against ultraviolet radiation (UVR), the main SC etiologic factor. Yet, the increasing SC incident have suggested that additional preventive agents could complement the sunscreen, helping to reverse this trend. Mutant p53 (mutp53) represents a promising target in SC prevention, since its occurrence is a key early event in UVR-induced SC. Thus, the inhibition of mutp53 formation by mutp53 reactivators, which are able to restore the wild-type-like function to p53, would be a valuable strategy for SC prevention. Recently, our group identified the tryptophanol-derived oxazoloisoindolinone SLMP53-2 as a new mutp53 reactivator. Herein, we aimed to study the potential of SLMP53-2 as a chemopreventive drug against UVR-induced SC. For that, assays with normal human keratinocyte HaCaT cells were performed. Cells were pre-treated with SLMP53-2 for 24h, followed by exposure to distinct UVB doses and further analysis. For in vivo studies, FVB/N mice were topically pre-treated for 1h prior to UVB radiation (180 mJ/cm2) and the skin was collected 24h after for immunohistochemistry/immunofluorescence analysis. Pre-treatment of HaCaT cells with SLMP53-2 enhanced cell survival in response to UVB with promotion of cell cycle arrest in G0/G1 phase, upregulation of p21 protein levels, reduction of UVB-induced apoptosis, and inhibition of JNK activity. SLMP53-2 also protected cells from UVB-induced ROS generation and from its consequent lipid peroxidation and protein carbonylation. It enhanced DNA repair through upregulation of the protein and mRNA levels of XPC (xeroderma pigmentosum complementation group C) and DDB-2 (DNA damaged binding protein 2) from the NER (nucleotide excision repair) pathway. In fact, SLMP53-2 depleted UVB-induced DNA damage, as evidenced by a reduction of DNA in comet tails, γH2AX staining and cyclobutane pyrimidine dimers (CPDs) levels. SLMP53-2 also showed promising activity in suppressing UVB-induced inflammation, decreasing COX-2, IL-6 and TNF-α protein levels and reducing NF-kB nuclear translocation and DNA binding ability. Moreover, SLMP53-2 enhanced the expression of key players in keratinocytes differentiation, namely NOTCH1 and keratin 1. Consistently, pre-treatment with SLMP53-2 reduced mutp53 protein levels by potentially restoring its wild-type p53 form, which may underlie the increased resistance of HaCaT cells to UVB harmful effects. Importantly, in vivo studies supported that SLMP53-2 pre-treatment of UVB-irradiated mice promoted the survival of epidermal cells, reducing the DNA damage by enhancement of DNA repair and downregulating mutp53 expression levels. Further, it reduced the expression of inflammatory-related proteins and promoted cell differentiation. Collectively, these results support a promising chemopreventive activity for SLMP53-2 against UVB-induced skin damage. We thank FCT/MCTES for the projects UIDB/50006/2020, UIDP/04423/2020 and fellowship SFRH/BD/128673/2017 (J.B. Loureiro).
Citation Format: Joana B. Loureiro, Rita Ribeiro, Nair Nazareth, Tiago Ferreira, Adelina Quaresma, Valentina Barcherini, Laura Marabini, Paula A. Oliveira, Maria M. M. Santos, Lucília Saraiva. Chemoprevention by the mutant p53 reactivator SLMP53-2 on ultraviolet radiation-induced skin cancer [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2021 Oct 7-10. Philadelphia (PA): AACR; Mol Cancer Ther 2021;20(12 Suppl):Abstract nr P030.