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Updated: Jun 4, 2026
Generation and Expansion of Primary, Malignant Pleural Mesothelioma Tumor Lines
Published on: April 21, 2022
Onconase mediated NFKβ downregulation in malignant pleural mesothelioma
C M Goparaju1, J D Blasberg, S Volinia
1New York University Medical Center, Department of Cardiothoracic Surgery, New York, NY 10016, USA.
Abstract:
Treatment of malignant pleural mesothelioma (MPM) with Ranpirnase (Onconase) results in disruption of protein translation and cell apoptosis. We hypothesize that Onconase exerts an effect via downregulation of nuclear factor kappa B (NFKβ) by specific microRNAs (miRNAs) and that interference of this pathway could have implications for MPM resistance to chemotherapy. Three immortalized MPM cell lines (H2959, H2373 and H2591) were exposed to Onconase at 0-20 microg/ml. Cell counts were measured at 48 and 72 h. Gene expression in miRNA-enriched RNA was validated by reverse transcription-PCR (RT-PCR). The functional implications of miRNA expression were evaluated by transfecting miRNA mimics or inhibitors into MPM cell lines, and performing Matrigel invasion, cell proliferation, soft agar colony formation and scratch closure assays. Effects on NFKβ expression and downstream targets including ABC transporters, BCL-xl and IAP were assessed by RT-PCR and western blotting. Treatment with 20 microg/ml of Onconase significantly decreased cell count and invasion. Hsa-miR-17* was significantly upregulated and hsa-miR-30c was significantly downregulated by Onconase treatment in all cell lines. Forced expression of hsa-miR-17* mimic and hsa-miR-30c inhibitor each significantly decreased functional activity of Onconase in all assays. NFKB1 (p50) expression and downstream targets were also decreased with Onconase treatment, as well as with forced expression of miRNA mimic and inhibitors. Onconase treatment caused a significant decrease in cell proliferation, invasion and in expression of certain miRNAs. Recapitulation of the resultant miRNA expression pattern with hsa-miR-17* mimic and hsa-miR-30c inhibitor resulted in downregulation of NFKB1 and reduced malignant behavior in functional assays. Thus, Onconase likely exerts its antitumor effect through these miRNAs.
Insights
Ranpirnase (Onconase) treats malignant pleural mesothelioma by altering microRNAs (miRNAs) and downregulating nuclear factor kappa B (NFKβ). This mechanism reduces cancer cell proliferation and invasion, offering potential for chemotherapy resistance.
Area of Science:
- Oncology
- Molecular Biology
- Gene Expression Regulation
Background:
- Malignant pleural mesothelioma (MPM) is a challenging cancer with limited treatment options.
- Ranpirnase (Onconase) is an investigational drug that disrupts protein translation and induces apoptosis in MPM cells.
- The precise molecular mechanisms by which Onconase exerts its anti-tumor effects, particularly its interaction with signaling pathways like NF-κB, require further elucidation.
Purpose of the Study:
- To investigate the hypothesis that Onconase downregulates nuclear factor kappa B (NF-κB) via specific microRNAs (miRNAs).
- To explore the role of this miRNA-NF-κB pathway in mediating Onconase's anti-cancer effects in MPM.
- To assess the potential implications of this pathway for overcoming chemotherapy resistance in MPM.
Main Methods:
- Exposure of three immortalized MPM cell lines (H2959, H2373, H2591) to varying concentrations of Onconase.
- Validation of miRNA expression changes (hsa-miR-17*, hsa-miR-30c) using RT-PCR.
- Functional assays (Matrigel invasion, proliferation, colony formation, scratch closure) following transfection with miRNA mimics or inhibitors.
- Assessment of NF-κB (NFKB1) expression and downstream targets (ABC transporters, BCL-xl, IAP) via RT-PCR and Western blotting.
Main Results:
- Onconase treatment significantly reduced MPM cell count and invasion.
- Onconase upregulated hsa-miR-17* and downregulated hsa-miR-30c in all tested cell lines.
- Forced expression of hsa-miR-17* mimic or hsa-miR-30c inhibitor mimicked Onconase's effects, reducing malignant behavior and downregulating NFKB1 expression.
Conclusions:
- Onconase likely exerts its anti-tumor effects in malignant pleural mesothelioma through the modulation of specific miRNAs (hsa-miR-17* and hsa-miR-30c).
- These miRNAs appear to mediate their effects by downregulating NF-κB signaling and its downstream targets.
- Understanding this miRNA-mediated pathway provides novel insights into Onconase's mechanism of action and potential therapeutic strategies for MPM.
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