Elevated expression of PAI-1 has been widely linked with adverse outcomes in a variety of human cancers, such as breast, gastric and ovarian cancers, rendering PAI-1 a prognostic biomarker. As a result, several chemical inhibitors are currently being developed against PAI-1; however, the clinical setting where they might confer survival benefits has not yet been elucidated.
RNA sequencing data analysis from the TCGA/GTEx cancer portals (n = 3607 samples). In silico molecular docking analyses to predict functional macromolecule interactions. ER-/PR- (MDA-MB-231) and ER+/PR+ (MCF-7) breast cancer cell lines implemented to assess the effect of oleuropein as a natural inhibitor of PAI-1-mediated oncogenic proliferation.
We show that high PAI-1 levels inversely correlate with ER and PR expressions in a wide panel of estrogen/progesterone-responsive human malignancies. By implementing an in silico molecular docking analysis, we identify oleuropein, a phenolic component of olive oil, as a potent PAI-1-binding molecule displaying increased affinity compared to the other olive oil constituents. We demonstrate that EVOO or oleuropein treatment alone may act as a natural PAI-1 inhibitor by incrementally destabilising PAI-1 levels selectively in ER-/PR- breast cancer cells, accompanied by downstream caspase activation and cell growth inhibition. In contrast, ER+/PR+ breast cancer cells, where PAI-1 expression is absent or low, do not adequately respond to treatment.
Our study demonstrates an inverse correlation between PAI-1 and ESR1/PGR levels, as well as overall patient survival in estrogen/progesterone-responsive human tumours. With a focus on breast cancer, our data identify oleuropein as a natural PAI-1 inhibitor and suggest that oleuropein-mediated PAI-1 destabilisation may confer clinical benefit only in ER-/PR- tumours.
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We would like to thank Yanni’s Olive Grove Company in Potidea Chalkidiki, Greece for providing the early harvest EVOO.
This research is co-financed by Greece and the European Union (European Social Fund- ESF) through the Operational Program «Human Resources Development, Education and Lifelong Learning» in the context of the project ‘Reinforcement of Postdoctoral Researchers - 2nd Cycle’ (grant number: 2019-050-0503-18066), implemented by the State Scholarships Foundation (ΙΚΥ).
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Supplementary Fig. 1. Densitometry providing quantification for Western blots. (a) Quantification of PAI-1 and CASP-8 levels based on β-actin expression for Fig. 3a. (b) Same as (a), for Fig. 3c. (c) Same as (a), for Fig. 3e. **P<0.01 and ***P<0.001, respectively, of Student’s t-test; n.s.: non-significant. Error bars indicate s.e.m. Data shown are representative of at least 3 independent experiments. Supplementary Fig. 2 PAI-1 and TP53 correlation in estrogen/progesterone-responsive tumours. (a) Correlation analysis between PAI-1 and TP53 levels expressed in transcripts per million (TPM) across 9 estrogen/progesterone-responsive human tumour types (BRCA, COAD, LUAD, LUSC, OV, PRAD, READ, UCEC, UCS; n indicates total number of tumour samples). RNA seq data were retrieved from the TCGA and GTEx databases and analysed using the GEPIA and GEPIA2 online tools. (b) Overall survival data of cancer patients in (a) based on TP53 expression levels. TP53 level changes do not confer a significant effect on survival (p=0.58). See also Fig 1. Data were retrieved from the TCGA and GTEx databases using the GEPIA online tool(pptx 335 kb)
Supplementary Table 1 Characterisation of the phenolic and non-phenolic content of EVOO used in in vitro experiments (pptx 73 kb)
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Tzekaki, E.E., Geromichalos, G., Lavrentiadou, S.N. et al. Oleuropein is a natural inhibitor of PAI-1-mediated proliferation in human ER-/PR- breast cancer cells. Breast Cancer Res Treat 186, 305–316 (2021). https://doi.org/10.1007/s10549-020-06054-x
- Plasminogen activator inhibitor-1 (PAI-1)
- Extra virgin olive oil (EVOO)
- ER/PR-responsive human cancer