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The p53 reactivator PRIMA-1MET synergises with 5-fluorouracil to induce apoptosis in pancreatic cancer cells

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Abstract

Pancreatic cancer (PC) is one of the deadliest malignancies; p53 is mutated in approximately 75% of PC patients. Hence, the protein derived from mutant/wild-type TP53 may represent a therapeutic target. Interestingly, a p53 reactivator (PRIMA-1MET) showed promise in clinical trials of haematological malignancies; therefore, it warrants an in vitro evaluation in PC cell lines. To evaluate the antiproliferative effects of PRIMA-1MET, either alone or combined with the common chemotherapy 5-fluorouracil (5-FU), against mutated and wild-type p53 PC cell lines. This study involved p53-mutant (AsPC-1) and p53-wild type (Capan-2) PC cell lines. The cytotoxicity of PRIMA-1MET alone or in combination with 5-FU was evaluated by MTT assay. Synergism was assessed by calculating the combination index (CI) via CalcuSyn software. Fluorescence microscopy was used to analyse apoptosis following acridine orange/ethidium bromide (AO/EB) staining. Morphological changes were investigated with an inverted microscope. Quantitative reverse transcription PCR (RT‒qPCR) was used to measure gene expression. Both PC cell lines were sensitive to PRIMA-1MET monotherapy. Furthermore, PRIMA-1MET and 5-FU had a synergistic effect (CI < 1), reflected by significant enhancement of apoptosis and morphological changes in the combination vs. monotherapy treatments. Moreover, the RT‒qPCR results indicated increased expression of the NOXA and TP73 genes in combination-treated cells. Our data suggested that PRIMA-1MET, whether alone or combined with 5-FU, has an antiproliferative effect on PC cell lines regardless of p53 mutational status. The synergism of the combination was associated with significant apoptosis induction through p53-dependent and p53-independent pathways. Preclinical confirmation of these data in in vivo models is highly recommended.

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Data Availability

The datasets generated during the current study are available from the corresponding author upon reasonable request.

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Acknowledgements

The authors thank Dr. Basma Talib Jasim for providing cell lines. We also thank Dr. Wrood Salim Dawood for her assistance in the TC lab of the College of Pharmacy, Mustansiriyah University. We also thank biologist Shahad Ali Abdulameer Mudhafar for offering help when needed for the work done in the Biotechnology Research Center, Al-Nahrain University labs.

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The authors funded this study, and no other funding was gained.

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Authors and Affiliations

  1. Department of Pharmacology and Toxicology, College of Pharmacy, Mustansiriyah University, Baghdad, Iraq

    Ibtehal Mohammed & Inam Sameh Arif

  2. Medical and Molecular Biotechnology Department, Biotechnology Research Center, Al-Nahrain University, Jadriya, Baghdad, Iraq

    Ali Haider Alhammer

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All authors contributed to the study conception, design, and material preparation. I.M. and A.A. performed data collection and analysis. A.A. wrote the first draft of the manuscript, and all authors commented on previous versions of the manuscript. All authors reviewed the manuscript.

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Correspondence to Ali Haider Alhammer.

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Mohammed, I., Alhammer, A.H. & Arif, I.S. The p53 reactivator PRIMA-1MET synergises with 5-fluorouracil to induce apoptosis in pancreatic cancer cells. Invest New Drugs 41, 587–595 (2023). https://doi.org/10.1007/s10637-023-01380-5

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