Evening Primrose Extract Modulates TYMS Expression via SP1 Transcription Factor in Malignant Pleural Mesothelioma

Our plain-English summary of Evening Primrose Extract Modulates TYMS Expression via SP1 Transcription Factor in Malignant Pleural Mesothelioma, by M. Chmielewska-Kassassir, K. Sobierajska, W.M. Ciszewski, et al (Cancers, 2023).

The study

• Authors: M. Chmielewska-Kassassir, K. Sobierajska, W.M. Ciszewski, J. Kryczka, A. Zieleniak, L.A. Wozniak
• Journal: Cancers
• Published: 2023;15(20)
• Cited by: 3 articles, as counted by NIH iCite
• Read it: PubMed · Publisher · Free full text

Abstract

Purpose: To determine the mechanism of EPE in downregulating TYMS in MPM cancer.

Methods: The TYMS mRNA expression with epithelial-to-mesenchymal transition biomarkers and nuclear factor SP1 was assessed using the GEO database in a data set of MPM patients (GSE51024). Invasive MPM cell lines were in vitro models for the investigation of TYMS expression after EPE treatment. The tyms promoter SP1 binding sequences were determined using Genomatix v 3.4 software Electrophoretic mobility shift and dual-luciferase reporter assays revealed specific SP1 motifs in the interaction of EPE and reference compounds. Chromatin immunoprecipitation and Re-ChIP were used for the co-occupancy study.

Results: In MPM patients, a positive correlation of overexpressed TYMS with mesenchymal TWIST1, FN1 and N-cadherin was observed. EPE and its major components, gallic and ellagic acid (GA and EA, respectively), downregulated TYMS in invasive MPM cells by interacting with particular SP1 motifs on the tyms promoter. The luciferase constructs confirmed the occupation of two SP1 regulatory regions critical for the promotion of TYMS expression. Both EPE and reference standards influenced SP1 translocation into the nucleus.

Conclusion: EPE components reduced TYMS expression by occupation of SP1 motifs on the tyms promoter and reversed the EMT phenotype of invasive MPM cells. Further in-depth analysis of the molecular docking of polyphenol compounds with SP1 regulatory motifs is required.

In plain English

This laboratory study looked at malignant pleural mesothelioma (MPM). It did not examine asbestos exposure. The authors first analysed a public gene-expression dataset of 96 samples: 54 tumor and 42 adjacent non-tumor tissue. TYMS, an enzyme linked to resistance to folate-based chemotherapy, was higher in tumors. Its levels correlated positively with the markers TWIST1, FN1 and N-cadherin. The authors then treated three MPM cell lines (JU77, MSTO-H211 and NCI-H28) with an extract of defatted evening primrose seeds (EPE). They also tested its main components, gallic acid and ellagic acid, and urolithin A, which gut microbes produce from ellagic acid. In the two more invasive lines, JU77 and MSTO-H211, EPE lowered TYMS and reduced the protein SP1 in the cell nucleus, by 37% in MSTO-H211 and 31% in JU77. It also shifted cell markers away from a mesenchymal (invasive) pattern toward an epithelial pattern. Single compounds had weaker effects, and none of these changes were seen in NCI-H28. The authors concluded that EPE components reduced TYMS through SP1 binding sites and reversed this invasive pattern. No people or animals received the extract.

We wrote this plain-English summary from the full article with the help of AI, and a second AI model checked each sentence against the article. It is not the authors' wording; the abstract and the article are their own account.

What the study did and found

• Design: Laboratory (in vitro) study of mesothelioma cell lines, combined with bioinformatic analysis of a public patient gene-expression dataset
• Who was studied: Public GEO dataset GSE51024 of 96 samples (54 surgically resected MPM tumor samples, 42 adjacent non-malignant tissue samples); three human MPM cell lines: sarcomatoid JU77, biphasic MSTO-H211 and pleural effusion metastatic NCI-H28
• Exposure: No asbestos exposure was studied. Cells were treated with evening primrose (Oenothera paradoxa) seed extract at 200 µg/mL (corresponding to 41.6 µg/mL gallic acid equivalents). They were also treated with single compounds: gallic acid at 10.3 µg/mL and ellagic acid at 15.4 µg/mL, or gallic acid, ellagic acid or urolithin A at 41.6 µg/mL. Urolithin A is described as an ellagic acid metabolite; the article's cited source calls urolithins gut microbial metabolites.
• Main results:
  • In the patient dataset, TYMS, TWIST1, N-cadherin (CDH-2) and FN1 were higher in tumors than in paired adjacent tissue. Vimentin, ZEB-1 and Snail showed no significant differences. E-cadherin (CDH-1) was lower in tumors.
  • In the patient dataset, TYMS correlated positively with TWIST, FN1 and CDH-2 but not with ZEB1 and VIM. SP1 showed no statistically significant correlation except with TWIST1 (0.309).
  • In untreated cells, TYMS mRNA was approximately 1.5 times higher in the more invasive JU77 and MSTO-H211 lines than in the low-invasive NCI-H28 line, and TYMS protein was 3.0-fold higher
  • Across the cell lines, the correlation coefficient (R) between invasion ability (taken from the authors' earlier study) and TYMS level was 0.84
  • EPE treatment significantly decreased TYMS expression in MSTO-H211 and JU77 cells but not in NCI-H28. SP1 levels were not changed in any cell line.
  • In JU77 and MSTO-H211 cells, EPE decreased EMT transcription factors and mesenchymal markers and increased epithelial markers. This was not detected in NCI-H28.
  • Single compounds (gallic acid, ellagic acid, urolithin A) lowered TYMS less than EPE did, at both concentrations tested
  • Compared with untreated cells, EPE reduced nuclear SP1 protein by 37% in MSTO-H211 and 31% in JU77 cells. Gallic acid and ellagic acid alone, at EPE-equivalent doses, gave reductions of 19% and 20%. At 41.6 µg/mL, ellagic acid, gallic acid and urolithin A gave reductions of 25%, 21% and 24%. There were no significant changes in NCI-H28.
  • In invasive MPM cells, EPE components inhibited SP1 binding to the region of the tyms promoter from position minus 241 to minus 21
• The authors' conclusion: The authors concluded that EPE components reduced TYMS expression by occupying SP1 motifs on the tyms promoter, and reversed the EMT phenotype of invasive MPM cells. They suggested that the whole polyphenol-rich extract has a benefit over individual compounds, and called for further molecular docking analysis.
• Limitations the authors state:
  • The authors noted that the heterogeneous patient material, and SP1's complex activity on many target genes, might complicate their results

Topics

Medical terms in this study, explained in our Lexicon: Pleural Mesothelioma, Biomarker, Translocation.

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