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P53 reactivator PRIMA-1 MET synergizes with 5-Fluorouracil to induce apoptosis in pancreatic tumor cells
  • Ibtehal Mohammed,
  • Ali Haider Alhammer,
  • Inam Sameh Arif
Ibtehal Mohammed
Mustansiriyah University College of Pharmacy
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Ali Haider Alhammer
Al-Nahrain University Biotechnology Research Center

Corresponding Author:[email protected]

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Inam Sameh Arif
Mustansiriyah University College of Pharmacy
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Abstract

Pancreatic cancer (PC) is one of top deadliest cancer types, p53 was found mutated in about 75% of PC patients. Hence, mutant/wild-type p53 protein may represent a unique therapeutic target. Interestingly, a p53-reactivator (PRIMA-1 MET) was recently approved by FDA and showed promise in hematological malignancies, therefore, warrants an in vitro evaluation in PC cell lines. The aim of this study was valuation of the antiproliferative effects of PRIMA-1 MET single or combined with an essential chemotherapy 5-fluorouracil (5-FU) in both mutated and wild-type p53 PC cell lines. In this study, p53-mutant (AsPC-1) and p53-wild type (Capan-2) PC cell lines were used. In vitro cytotoxicity of PRIMA-1 MET was evaluated single or combined with 5-FU by MTT assay. The Chou and Talalay approach was employed to evaluate synergism by calculating the combination index (CI) by CalcuSyn software. Apoptosis was analyzed by fluorescent microscopy following Acridine orange/ethidium bromide (AO/EB) staining. Morphological changes were investigated by inverted microscope. Real-time quantitative PCR (RT-qPCR) was used to measure gene expression. Both PC cell lines (p53-wt and p53-mut) have shown sensitivity to PRIMA-1 MET monotherapy. Furthermore, adding PRIMA-1 MET to 5-FU was synergistic (CI<1) and this enhanced effect was reflected in greater induction of apoptosis and morphological changes. Moreover, RT-qPCR results indicated increased expression of Noxa and TP73 genes in combination treated cells. Our data suggested that PRIMA-1 MET, whether single or combined with 5-FU, have an antiproliferative impact on PC cell lines regardless of p53 mutational status. Synergism was allied with significant apoptosis induction through p53-dependent and p53-independent pathways. Preclinical confirmation of these data in in vivo models is highly recommended.
04 Jul 2023Published in Investigational New Drugs. 10.1007/s10637-023-01380-5