Protective Effect of Coriolus versicolor Cultivated in Citrus Extract Against Nitric Oxide-Induced Apoptosis in Human Neuroblastoma SK-N-MC Cells

Abstract

Nitric oxide (NO) is a reactive free radical and a messenger molecule in many physiological functions. However, excessive NO is believed to be a mediator of neurotoxicity. The medicinal plant Coriolus versicolor is known to possess anti-tumor and immune-potentiating activities. In this study, we investigated whether Coriolus versicolor possesses a protective effect against NO donor sodium nitroprusside (SNP)-induced apoptosis in the human neuroblastoma cell line SK-N-MC. We utilized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, flow cytometry, 4,6-diamidino-2-phenylindole (DAPI) staining, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, DNA fragmentation assay, reverse transcription-polymerase chain reaction (RT-PCR), Western blot analysis, and caspase-3 enzyme activity assay in SK-N-MC cells. MTT assay showed that SNP treatment significantly reduces the viability of cells, and the viabilities of cells pre-treated with the aqueous extract of Coriolus versicolor cultivated in citrus extract (CVEcitrus) was increased. However, aqueous extract of Coriolus versicolor cultivated in synthetic medium (CVEsynthetic) showed no protective effect and aqueous citrus extract (CE) had a little protective effect. The cell treated with SNP exhibited several apoptotic features, while those pre-treated for 1 h with CVEcitrus prior to SNP expose showed reduced apoptotic features. The cells pre-treated for 1 h with CVEcitrus prior to SNP expose inhibited p53 and Bax expressions and caspase-3 enzyme activity up-regulated by SNP. We showed that CVEcitrus exerts a protective effect against SNP-induced apoptosis in SK-N-MC cells. Our study suggests that CVEcitrus has therapeutic value in the treatment of a variety of NO-induced brain diseases.

Nitric Oxide as a Potential Adjuvant Therapeutic for Neuroblastoma: Effects of NO on Murine N2a Cells

“…This data further corroborates the previous claims: GSNO-treated cells moderately decrease in cellular viability and the observed decrease is due to NO-release from GSNO. Again, this result was in accordance with similar NO-based anticancer therapeutic studies [ 21 , 24 ] and demonstrates the potential of NO-based anticancer adjuvants.…”

“…Ultimately, the observed cellular viability of N2a cells treated with GSNO assessed using each assay was remarkably similar, providing important evidence of the antitumor potential of NO in neuroblastoma treatment. The data observed here further corroborated the utility of NO-based anticancer therapeutics that has been presented in various other studies [21,24]. Clearly, the level of reduction identified in this study does not warrant a stand-alone anticancer treatment; however, NO has the potential to be exploited for use as an adjuvant.…”

“…It is important to reiterate that treated GSH samples were found to be 100% ± 5% viable, indicating that NO was the active therapeutic, not GSNO. Studies done by Kim et al as well as Suchyta and Schoenfisch on NO-based anticancer therapeutics have shown analogous results, indicating reductions in cellular viability in up to 60% of human SK-N-MC neuroblastoma cells and up to 66% in ovarian cancer cell lines, respectively [21,24]. Although moderate, the effect observed here is particularly promising as it demonstrates the potential of NO-releasing therapeutics to serve as adjuvants to other anticancer therapeutic approaches while decreasing detrimental side effects to the patient.…”

“…The multitude of new therapies include cytotoxic agents, targeted agents, immunotherapy, retinoids, angiogenesis inhibitors, tyrosine kinase inhibitors, as well as other approaches [ 12 ]. One approach that has been of little focus in pediatric neuroblastoma research is the use of nitric oxide (NO) as an anticancer agent [ 20 , 21 ]. Numerous studies have shown that NO effectively induces tumor-specific cytotoxicity and pharmaceutical applications of S -Nitrosylation have exhibited great potential for use as adjuvant therapeutics in the clinical management of cancer [ 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 ].…”

RNA-binding protein RBM3 prevents NO-induced apoptosis in human neuroblastoma cells by modulating p38 signaling and miR-143

“…The NO donor SNP is a well-established toxin that can trigger apoptosis in cultured neurons and neuroblstoma cells 6 12 27 28 29 30 31 32 33 . To determine whether cooling protects neural cells from NO-induced apoptosis, human SH-SY5Y neuroblastoma cells were treated with various concentrations of SNP.…”