Oncogenic mutation p53 Y220C is the ninth most common mutation of p53 protein. The Y220C mutation creates an expanded surface pocket in the DNA-binding domain, rapidly unfolds and denatures under physiological conditions, which effectively cancels p53 signaling and leads to the development of a tumor. In this work the novel indazole derived compounds were tested. It was found that these molecules bind to the mutated p53 protein and stabilize its structure in vitro.
In this work the human breast carcinoma (MCF7 p53 wt, MCF7 p53 Y220C, MCF7 p53 -/-) and the human hepatocarcinoma (HUH7 p53 Y220C) cell lines were used. Cell viability was evaluated using colorimetric MTS test. The cells were treated with compounds at the concentration range of 0-200 μM. The obtained data were processed in GraphPad Prism. The expression of p53 target genes upon treatment with the compounds was also quantified using real-time quantitative PCR (RT-qPCR).
Viability of cells incubated with the compounds were determined for cell lines with different p53 status. IC50 values for cells with wild-type p53 status (MCF7 p53 wt) and cells with TP53 knockout (MCF7 p53 -/-) could not be detected in the selected concentration range while in cells with mutated p53 (HUH7 p53 Y220C and MCF7 p53 Y220C) IC50 values were determined in the range of 32-55 μM. Expression of PUMA (BBC3) was found to be upregulated in HUH7 p53 Y220C and MCF7 p53 Y220C cells upon treatment with the compounds.
The results demonstrated that the compounds exert targeted effect on cells carrying p53 Y220C mutation. We established that the compounds selectively reduced the viability of p53 Y220C cell lines and upregulated transcription of p53 target genes associated with apoptosis in p53 Y220C-dependent manner. The work was funded by grant from the Russian Science Foundation 22-24-20034 and supported by the Kazan Federal University Strategic Academic Leadership Program (PRIORITY-2030).
Kazan Federal University.
The work was funded by grant from the Russian Science Foundation 22-24-20034 and supported by the Kazan Federal University Strategic Academic Leadership Program (PRIORITY-2030).
All authors have declared no conflicts of interest.