Induction of apoptosis through extrinsic/intrinsic pathways and suppression of ERK/NF‐κB signalling participate in anti‐glioblastoma of imipramine

Abstract

Glioblastomas are the most aggressive type of brain tumour, with poor prognosis even after standard treatment such as surgical resection, temozolomide and radiation therapy. The overexpression of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) in glioblastomas is recognized as an important treatment target. Thus, an urgent need regarding glioblastomas is the development of a new, suitable agent that may show potential for the inhibition of extracellular signal-regulated kinase (ERK)/NF-κB-mediated glioblastoma progression. Imipramine, a tricyclic antidepressant, has anti-inflammatory actions against inflamed glial cells; additionally, imipramine can induce glioblastoma toxicity via the activation of autophagy. However, whether imipramine can suppress glioblastoma progression via the induction of apoptosis and blockage of ERK/NF-κB signalling remains unclear. The main purpose of this study was to investigate the effects of imipramine on apoptotic signalling and ERK/NF-κB-mediated glioblastoma progression by using cell proliferation (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide [MTT] assay), flow cytometry, Western blotting, and cell invasion/migration assay analysis in vitro. The ERK and NF-κB inhibitory capacity of imipramine is detected by NF-κB reporter gene assay and Western blotting. Additionally, a glioblastoma-bearing animal model was used to validate the therapeutic efficacy and general toxicity of imipramine. Our results demonstrated that imipramine successfully triggered apoptosis through extrinsic/intrinsic pathways and suppressed the invasion/migration ability of glioblastoma cells. Furthermore, imipramine effectively suppressed glioblastoma progression in vivo via the inhibition of the ERK/NF-κB pathway. In summary, imipramine is a potential anti-glioblastoma drug which induces apoptosis and has the capacity to inhibit ERK/NF-κB signalling. K E Y W O R D S ERK, extrinsic/intrinsic apoptosis, glioblastoma, imipramine, NF-κB | 3983 HSU et al.

Identification of a Metabolism-Related Risk Signature Associated With Clinical Prognosis in Glioblastoma Using Integrated Bioinformatic Analysis

“…Interestingly, our results suggest that some conventional antipsychotics among drugs mentioned above, including amitriptyline, fluoxetine, and imipramine, may also play a surprising role in the treatment of GBM, which is consistent with previous studies and offers new hope for patients with GBM (12,75,76,80,83,84,104). In addition, Leite et al (105) demonstrated that clomipramine (tricyclic antidepressant drugs such as imipramine) has an impact on GBM growth and has no toxicity in normal cells (astrocytes and microglia).…”

“…In this study, we used CMap analysis to accurately identify compounds that have been shown to have specific effects on GBM or other tumor types by comparing the different expression genes of GMB samples from 3 clusters. These compounds include the DNA synthesis inhibitor anisomycin ( 73 ), glutamate receptor antagonist amantadine ( 74 ), norepinephrine inhibitor amitriptyline ( 75 , 76 ), solute carrier family member inhibitor bumetanide ( 77 ), PPAR receptor agonist clofibrate ( 78 ), and fenofibrate ( 79 ), selective serotonin reuptake inhibitor (SSRI) fluoxetine ( 75 , 80 ), ATPase inhibitor helveticoside ( 81 , 82 ), norepinephrine reuptake inhibitor imipramine ( 75 , 76 , 83 , 84 ), opioid receptor agonist loperamide ( 85 ), phosphodiesterase inhibitor papaverine ( 86 , 87 ) and rolipram ( 88 – 92 ), polar auxin transport inhibitor quercetin ( 93 , 94 ), cyclooxygenase inhibitor rofecoxib ( 95 , 96 ), calcineurin inhibitor tacrolimus ( 97 ), purinergic receptor antagonist ticlopidine ( 98 , 99 ), HDAC inhibitor vorinostat ( 100 ), Rho associated kinase inhibitor Y-27632 ( 101 – 103 ).…”

Induction of Apoptosis, Inhibition of MCL-1, and VEGF-A Expression Are Associated with the Anti-Cancer Efficacy of Magnolol Combined with Regorafenib in Hepatocellular Carcinoma

“…The Hep3B and SK-Hep1 cells (2 × 10 5 ) were seeded in a 10-cm dish and incubated with 50 μM magnolol, 10 μM regorafenib, and a combination for 48 h. Forty micrograms of protein were separated by 8–12% SDS-PAGE and transferred onto a PVDF membrane [ 15 , 16 ]. Primary antibodies against CyclinD1, X-linked inhibitor of apoptosis (XIAP), cellular FLICE (FADD-like IL-1β-converting enzyme)-inhibitory protein (c-FLIP), MCL-1, matrix metallopeptidase 9 (MMP9), VEGF-A, mediator of DNA damage checkpoint 1 (MDC-1), Bcl-2-associated X protein (BAX), Bcl-2 homologous antagonist killer (BAK), FAS, FAS-L, cleaved-caspase-3, -8, and -9, cleaved Poly [ADP-ribose] polymerase 1 (PARP-1), and β-actin (Elabscience) were used for the conjugation overnight, followed by incubation with secondary antibodies.…”

“…For subG1 analysis, the harvested cells were fixed by 70% ethanol overnight at −20 °C and stained by a PI/RNase solution (cat: 550625, BD Biosciences). The fluoresce signal from the cells was detected and quantified by NovoCyte flow cytometry and the NovoExpress ® software (Agilent Technologies Inc., Santa Clara, CA, USA) [ 15 , 17 ].…”

Anticancer Efficacy and Mechanism of Amentoflavone for Sensitizing Oral Squamous Cell Carcinoma to Cisplatin

“…For annexin-V/PI analysis, cells were harvested and stained with Annexin-V/FTC apoptosis detection kit (Vazyme Biotech Co. Lt, Nanjing City, PR China) in the dark for 30 min at 37˚C. After different staining procedures, apoptosis-related signals in cells were finally evaluated by NovoCyte flow cytometry and quantified by NovoExpress ® software (16).…”