Min You1, Javier Varona-Santos1, Samer Singh2, David J Robbins2, Niramol Savaraj3, Dao M Nguyen4. 1. Thoracic Surgery Section, Division of Cardiothoracic Surgery, Department of Surgery, Sylvester Comprehensive Cancer Center, Leonard M. Miller School of Medicine, University of Miami, Miami, Fla. 2. Molecular Oncology Program, Department of Surgery, Sylvester Comprehensive Cancer Center, Leonard M. Miller School of Medicine, University of Miami, Miami, Fla. 3. Division of Oncology, Department of Medicine, Miami Veterans Affairs Medical Center, Sylvester Comprehensive Cancer Center, Leonard M. Miller School of Medicine, University of Miami, Miami, Fla. 4. Thoracic Surgery Section, Division of Cardiothoracic Surgery, Department of Surgery, Sylvester Comprehensive Cancer Center, Leonard M. Miller School of Medicine, University of Miami, Miami, Fla. Electronic address: dnguyen4@med.miami.edu.
Abstract
OBJECTIVE: The present study sought to determine whether the Hedgehog (Hh) pathway is active and regulates the cell growth of cultured malignant pleural mesothelioma (MPM) cells and to evaluate the efficacy of pathway blockade using smoothened (SMO) antagonists (SMO inhibitor GDC-0449 or the antifungal drug itraconazole [ITRA]) or Gli inhibitors (GANT61 or the antileukemia drug arsenic trioxide [ATO]) in suppressing MPM viability. METHODS: Selective knockdown of SMO to inhibit Hh signaling was achieved by small interfering RNA in 3 representative MPM cells. The growth inhibitory effect of GDC-0449, ITRA, GANT61, and ATO was evaluated in 8 MPM lines, with cell viability quantified using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cell death was determined by annexinV/propidium iodide staining and flow cytometry. RESULTS: SMO small interfering RNA mediated a two- to more than fivefold reduction of SMO and Gli1 gene expression as determined by real-time quantitative reverse-transcriptase polymerase chain reaction, indicating significant Hh pathway blockade. This was associated with significantly reduced cell viability (34% ± 7% to 61% ± 14% of nontarget small interfering RNA controls; P = .0024 to P = .043). Treating MPM cells with Hh inhibitors resulted in a 1.5- to 4-fold reduction of Gli1 expression. These 4 Hh antagonists strongly suppressed MPM cell viability. More importantly, ITRA, ATO, GANT61 induced significant apoptosis in the representative MPM cells. CONCLUSIONS: Hh signaling is active in MPM and regulates cell viability. ATO and ITRA were as effective as the prototypic SMO inhibitor GDC-0449 and the Gli inhibitor GANT61 in suppressing Hh signaling in MPM cells. Pharmaceutical agents Food and Drug Administration-approved for other indications but recently found to have anti-Hh activity, such as ATO or ITRA, could be repurposed to treat MPM.
OBJECTIVE: The present study sought to determine whether the Hedgehog (Hh) pathway is active and regulates the cell growth of cultured malignant pleural mesothelioma (MPM) cells and to evaluate the efficacy of pathway blockade using smoothened (SMO) antagonists (SMO inhibitor GDC-0449 or the antifungal drug itraconazole [ITRA]) or Gli inhibitors (GANT61 or the antileukemia drug arsenic trioxide [ATO]) in suppressing MPM viability. METHODS: Selective knockdown of SMO to inhibit Hh signaling was achieved by small interfering RNA in 3 representative MPM cells. The growth inhibitory effect of GDC-0449, ITRA, GANT61, and ATO was evaluated in 8 MPM lines, with cell viability quantified using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cell death was determined by annexinV/propidium iodide staining and flow cytometry. RESULTS:SMO small interfering RNA mediated a two- to more than fivefold reduction of SMO and Gli1 gene expression as determined by real-time quantitative reverse-transcriptase polymerase chain reaction, indicating significant Hh pathway blockade. This was associated with significantly reduced cell viability (34% ± 7% to 61% ± 14% of nontarget small interfering RNA controls; P = .0024 to P = .043). Treating MPM cells with Hh inhibitors resulted in a 1.5- to 4-fold reduction of Gli1 expression. These 4 Hh antagonists strongly suppressed MPM cell viability. More importantly, ITRA, ATO, GANT61 induced significant apoptosis in the representative MPM cells. CONCLUSIONS: Hh signaling is active in MPM and regulates cell viability. ATO and ITRA were as effective as the prototypic SMO inhibitor GDC-0449 and the Gli inhibitor GANT61 in suppressing Hh signaling in MPM cells. Pharmaceutical agents Food and Drug Administration-approved for other indications but recently found to have anti-Hh activity, such as ATO or ITRA, could be repurposed to treat MPM.
Authors: Asmaa Waheed Mohamed; Mohamed Elbassiouny; Dalia Abdelghany Elkhodary; May Ahmed Shawki; Amr Shafik Saad Journal: Med Oncol Date: 2021-02-09 Impact factor: 3.064
Authors: Bin Li; Camilla Giambelli; Bo Tang; Emily Winterbottom; Jun Long; Ke Jin; Zhiqiang Wang; Dennis Liang Fei; Dao M Nguyen; Mohammad Athar; Baolin Wang; Pochi R Subbarayan; Lily Wang; Priyamvada Rai; Bach Ardalan; Anthony J Capobianco; David J Robbins Journal: Mol Pharmacol Date: 2015-11-16 Impact factor: 4.436