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. 2022 Jan 24;12:1248. doi: 10.1038/s41598-022-05193-z

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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In vitro synergistic effects of MDM2 and MEK inhibition in CRC and PTC. (A) TP53 and MAPK pathway gene alteration frequencies in human CRCs and PTCs. (B,C) Three CRC cell lines (LoVo, HCT116, and B1011CL) and one PTC cell line (PDX.008CL) were treated for 72 h with increasing concentrations of vehicle, selumetinib, KRT-232, or a combination of both. Drug toxicity and cell growth were determined using sulforhodamine B assay. Individual IC₅₀ values (B) and combination indices (C) are shown. KRT-232 IC₅₀ was beyond the tested drug concentration range. CI < 1 indicates synergistic activity. IC₅₀ and CI values were calculated using CalcuSyn. (D) Colony formation assays of three CRC and one PTC cell line. Cells were treated with vehicle or various doses of selumetinib, KRT-232, or their combination for 2–3 weeks. This was followed by fixation of colonies and staining using crystal violet. (E) Individual IC₅₀ values were calculated using CalcuSyn. (F) The Bliss independence model was used to evaluate the drug interaction across all dose combinations tested. Interaction index values < 1 indicate synergy and values within 0–0.9 range were highlighted in green. All experiments were repeated at least three times.