Butein-instigated miR-186-5p-dependent modulation of TWIST1 affects resistance to cisplatin and bioenergetics of Malignant Pleural Mesothelioma cells

Our plain-English summary of Butein-instigated miR-186-5p-dependent modulation of TWIST1 affects resistance to cisplatin and bioenergetics of Malignant Pleural Mesothelioma cells, by M. Cioce, D. Rutigliano, A. Puglielli, et al (Cancer Drug Resistance, 2022).

The study

• Authors: M. Cioce, D. Rutigliano, A. Puglielli, V.M. Fazio
• Journal: Cancer Drug Resistance
• Published: 2022;5(3):814-828
• Cited by: 11 articles, as counted by NIH iCite
• Read it: PubMed · Publisher · Free full text

Abstract

Aim: Malignant pleural mesothelioma is a chemoresistant tumor, and biphasic and sarcomatoid histologies portend the worst prognosis for malignant pleural mesothelioma (MPM) patients. We obtained the microRNA expression profile of three biphasic-sarcomatoid MPM cell lines to identify commonly expressed microRNAs and evaluate the effect of butein, a chemo-sensitizing compound, on this microRNA subset. Methods: Nanostring-based microRNA profiling and analysis through the ROSALIND platform were employed to identify the commonly modulated microRNAs and their targets. MicroRNA-mimic transfection, Luciferase assay, and Western blotting were employed to show specific perturbation of TWIST1 levels by miR-186-5p. Sphere-forming assays, invasion assay, and metabolic profiling were used to assess the biological consequences of the butein-instigated miR-186-5p-mediated perturbation of TWIST1 levels. TGCA analysis was used to search for the correlation between TWIST1 and miR-186-5p levels in biphasic and epithelioid MPM specimens. Results: We identified a set of perturbed microRNAs, common to three biphasic/sarcomatoid MPM cell lines, after butein treatment. When focusing on miR-186-5p, we unraveled a butein-ignited and miR-186-5p-mediated modulation of TWIST1 levels which affected the 3D anchorage-independent growth, cisplatin resistance, invasion, and bioenergetics of the MPM cell lines tested. We showed that miR-186-5p and TWIST1 levels are anti-correlated in biphasic MPM specimens from TCGA. Conclusion: We unraveled a novel mechanism of action of butein, which attenuated the pro-tumorigenic features of MPM at least through a miR-186-5p-TWIST1 axis. We suggest that those activities converge into the chemo-sensitizing effect of this compound and may be of translational relevance.

© The Author(s) 2022.

In plain English

This was a laboratory study of three human malignant pleural mesothelioma (MPM) cell lines, plus an analysis of TCGA database tumor data. The cell lines were NCI-H2373, of sarcomatoid origin, and HP1 and MSTO-211H, of biphasic origin. The study did not examine asbestos exposure. The authors treated the cells with butein, a naturally occurring compound, at 10 µM for 8 hours. They reported 33 microRNAs (small RNA molecules that regulate genes) changed by butein, mostly downward. One that rose, miR-186-5p, lowered activity of a test gene carrying part of TWIST1, and both butein and miR-186-5p lowered TWIST1 protein. In lab tests, butein and miR-186-5p broke up tumor-cell clusters, reduced their resistance to the chemotherapy drug cisplatin and reduced glycolysis (sugar use). Butein also reduced invasion. Adding extra TWIST1 had opposite effects. In the TCGA data, miR-186-5p and TWIST1 levels were not significantly correlated across MPMs (P = 0.0748). Biphasic tumors (21) showed only a trend toward lower miR-186-5p and higher TWIST1 than epithelioid tumors (58). The authors concluded that butein acted at least partly through miR-186-5p and TWIST1. They noted they used no primary patient tumor samples.

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, with an added analysis of a public tumor database (TCGA)
• Who was studied: Three human malignant pleural mesothelioma cell lines: HP1 and MSTO-211H (biphasic origin) and NCI-H2373 (sarcomatoid origin). HEK293 cells were used as a non-MPM technical control. The database analysis used TCGA MPM specimens: 21 biphasic and 58 epithelioid.
• Exposure: Not asbestos. The cells were treated with butein (10 µM for 8 h) or with miR-186-5p mimics, or were made to overexpress TWIST1, and were then tested with the chemotherapy drug cisplatin.
• Main results:
  • Butein changed the levels of 33 microRNAs common to the three cell lines. Most of the changes were downward.
  • In MPM cells, miR-186-5p mimics lowered the activity of a luciferase reporter carrying the TWIST1 3' UTR. There was no significant change when the binding site was mutated.
  • Butein and miR-186-5p mimics caused NCI-H2373 spheroids to break apart, more so with cisplatin. TWIST1 overexpression produced compact spheroids that were resistant to cisplatin. The authors reported similar effects in MSTO-211H and HP-1 spheroids.
  • Butein strongly reduced invasion by NCI-H2373 cells, and TWIST1 overexpression increased it. HP1 cells did not invade at steady state and were not tested further.
  • In NCI-H2373 cells, butein and miR-186-5p mimics reduced glycolytic flux and oxygen consumption. Butein reduced oxygen consumption more. TWIST1 overexpression increased glycolytic flux but did not significantly change oxygen consumption (P = 0.07).
  • In TCGA MPMs there was no significant correlation between miR-186-5p and TWIST1 levels (Spearman rho = -0.1936, P = 0.0748). Biphasic tumors (n = 21) showed a trend toward lower miR-186-5p (P between 0.05 and 0.10) and higher TWIST1 than epithelioid tumors (n = 58).
• The authors' conclusion: The authors concluded that butein reduced pro-tumor features of MPM cells at least partly through a miR-186-5p and TWIST1 pathway. They suggested this contributes to butein's chemo-sensitizing effect and may be of translational relevance.
• Limitations the authors state:
  • The authors did not investigate whether butein's effect on miR-186-5p occurs in specific cell subpopulations.
  • The authors did not examine the mechanism by which lowering TWIST1 affects the pro-tumor program.
  • No primary MPM patient specimens were used in the experiments.
  • The metabolic tests were limited to two measures, ECAR and OCR.
  • How butein raises miR-186-5p was not resolved.

Topics

Medical terms in this study, explained in our Lexicon: Cisplatin, Pleural Mesothelioma, Biphasic Mesothelioma, Sarcomatoid, Prognosis, Epithelioid.

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