Silencing E3 Ubiqutin ligase ITCH as a potential therapy to enhance chemotherapy efficacy in p53 mutant neuroblastoma cells

Abstract

Methods Ethics approval. All in vivo procedures were approved by UCL animal care policies and were carried out under Home Office Licenses issued in accordance with the United Kingdom Animals (Scientific Procedures) Act 1986 (UK). Materials. 1,2-di-O-octadecenyl-3-trimethylammonium propane (DOTMA), 1,2-dipalmitoyl-sn-glycero-3-p hosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DPPE-PEG2000) and 1,2-dioleoyl-sn-glycero-3-p hosphoethanolamine (DOPE) were purchased from Avanti Polar Lipids, Inc. (Alabaster, AL, USA). Peptide ME27 (K16RVRRGACRGDCLG) was synthesized by Alta Bioscience (Birmingham, UK). Cell culture. Two p53-mutant neuroblastoma cell lines, Kelly 47,56 and SK-N-BE-2 (BE2) cells 5 were maintained in vitro in RPMI-1640 medium (Sigma, Dorset, UK) supplemented with 2 mM Glutamine and 10% FBS (Thermo Fisher, Paisley, UK). Cells were split every 3-4 days.

MHC Class I Downregulation in Cancer: Underlying Mechanisms and Potential Targets for Cancer Immunotherapy

“…Knockout of either of these proteins results in inadequate NFkB inhibition, indicating that all three proteins are indispensable in the functioning of this ubiquitin-editing complex [74]. Itch has been shown to be upregulated in several types of cancer, such as breast cancer and neuroblastoma, in which it was shown to play a major role in cancer progression [75][76][77][78][79]. The antidepressant clomipramine, its structural homologue norclomipramine, and 1,4-naphthoquinone 10E have been shown to reduce tumor growth and enhance chemotherapy in multiple cancer cell lines, including breast, prostate, and bladder cancer lines, as well as in a multiple myeloma xenograft model [80,81].…”

Deciphering the Role of p53 and TAp73 in Neuroblastoma: From Pathogenesis to Treatment

“…Similarly, RNA interference-mediated downregulation of Itch significantly enhanced suppression of pancreatic cancer growth by gemcitabine in vivo [ 97 ]. This was further demonstrated in NB cells by Meng et al, who used in vivo nano-delivery of the Itch siRNA in NB xenograft mouse models to sensitize tumor cells to radiotherapy [ 98 ]. Itch is responsible for the regulation of the proteasomal-dependent degradation of a group of target proteins, including TAp73 [ 99 ] ( Figure 1 b).…”

Non-Viral Carriers for Nucleic Acids Delivery: Fundamentals and Current Applications

“…Integrin binding was proposed for neuroblastoma cells. The peptide ME27 (RVRRGACRGDCLG) was shown to be able to initialize the αvβ3and αvβ5-targeted delivery into neuroblastoma in vivo [161]. Tet1 (HLNIL-STLWKYR) peptide is highly used for sphingomyelin and ganglioside GT1B and neuron targeting [156,162].…”