NF-κB-dependent mechanism of action of c-Myc inhibitor 10058-F4: Highlighting a promising effect of c-Myc inhibition in Leukemia cells, irrespective of p53 status

Due to the frequent contribution in the pathogenesis of different human malignancies, c-Myc is among those transcription factors that are believed to be pharmacologically targeted for cancer therapeutic approaches. In the present study, we examined the anti-leukemic effect of a well-known c-Myc inhibitor 10058-F4 on a panel of hematologic malignant cells harboring either mutant or wild-type p53. Notably, we found that the suppression of c-Myc was coupled with the reduction in the survival of all the tested leukemic cells; however, as far as we are aware, this study suggests for the first time that the cytotoxic effect of 10058-F4 was not significantly affected by the molecular status of p53. Delving into the molecular mechanisms of the inhibitor in the most sensitive cell line revealed that 10058-F4 could induce apoptotic cell death in mutant p53-expressing NB4 cells through the suppression of NF-κB pathway coupled with a significant induction of intracellular reactive oxygen species (ROS). In addition, we found that the anti-leukemic effect of 10058-F4 was overshadowed, at least partially, through the compensatory activation of the PI3K signaling pathway; highlighting a plausible attenuating role of this axis on 10058-F4 cytotoxicity. In conclusion, the results of the present study shed light on the favorable anti-leukemic effect of 10058-F4, especially in combination with PI3K inhibitors in acute promyelocytic leukemia; however, further investigations should be accomplished to determine the efficacy of the inhibitor, either as a single agent or in a combined-modal strategy, in leukemia treatment. © 2020, Iranian Journal of Pharmaceutical Research. All rights reserved

Contribution value of akt, c-myc, cip2a, and pp2a genes expression in leukemogenesis: A bright perspective on the molecular pattern of patients with acute myeloid leukemia (aml)

Background: The heterogeneous nature of acute myeloid leukemia (AML) and the hurdle to find a suitable treatment strategy for this malignancy put this type of leukemia at the top of the list of the priorities for finding a valuable biomarker to improve its treatment and predict the outcome of the patients. Objectives: Given the involvement of the variety of signaling pathways, foremost the PI3K axis in the pathogenesis of human can-cers, we aimed to investigate the expression of the most important downstream targets of this pathway to propose a plausible mechanism underlying AML pathogenesis. Methods: In this case-control study, the blood samples from 30 patients diagnosed with AML were collected and after extracting their RNAs, the expression levels of Akt, c-Myc, CIP2A, and PP2A were evaluated using qRT-PCR analysis. For the control group, we also collected blood samples from 10 healthy volunteers. Afterward, by applying statistical analysis, we determined the probable correlation between the expressions of the aforementioned genes. Results: There was a significant elevation in the expression levels of Akt, c-Myc, and CIP2A coupled with the meaningful reduction in the expression level of PP2A in AML samples. However, we failed to find any significant association between the expression level of the indicated genes and age, sex, and the percentage of the blasts. Conclusions: As the most straightforward interpretation of our results, we propose that probably the association between PI3K and c-Myc which is built through the interaction between CIP2A and PP2A may play a pivotal role in the pathogenies of AML and any component of this axis could serve as a potential new target for more profound treatment strategy. However, further detailed inves-tigations in this field are required to clarify the exact role of this interesting testis-specific pathway in the context of hematological malignancies, in particular AML. © 2020, Author(s)

Anti-Cancer Effect of Aprepitant on Nb4 Leukemic Cells

BACKGROUND AND OBJECTIVE: Genetic studies have demonstrated that the neurokini-1 Receptor (NK1R (is frequently involved in the pathogenesis of wide assortment of human malignancies, including acute promyelocytic leukemia (APL). The activity of this pathway in leukemic cells results in an excessive cell proliferation and evade from apoptosis. In this study, we aimed to investigate the effect of Aprepitant (NK1R antagonist) on the survival rate of APL cells. METHODS: This experimental study is conducted on APL-derived NB4 cells (Institute Pasteur). To determine the anti-tumor effect of Aprepitant, NB4 cells were divided into 6 groups: control and 1-, 2-, 3-, 4- and 5 µM-drug treated groups. Then the cell viability, metabolic activity, induction of apoptosis and transcriptional alteration of Bax and Bcl-2 genes were investigated after 24 and 36 h treatment using trypan blue assay, MTT assay, Annexin-V/PI staining and RQ-PCR analysis, respectively. FINDINGS: 36 h treatment with the highest concentration of Aprepitant (5 µM) resulted in an approximately 50% reduction in the viability (assessed by trypan blue) and metabolic activity (assessed by MTT assay) of NB4 cells (p<0.001) in comparison with control group. Moreover, Aprepitant is able to increase the proportion apoptotic cells from 1.4% in control group to 10.6% in 5 µM drug-treated cells though up-regulating Bax/Bcl-2 molecular ratio (p≤0.05). CONCLUSION: Aprepitant exerted both cytotoxic and anti-proliferative effects in NB4 cells

c-Myc Inhibition Using 10058-F4 Increased the Sensitivity of Acute Promyelocytic Leukemia Cells to Arsenic Trioxide Via Blunting PI3K/NF-κB Axis

Backgrounds: Although ATO is widely used to treat acute promelocytic leukemia (APL), the appropriate effects of the drug as a single agent are achieved in high doses which are not clinically achievable without the risk of side effects; highlighting the necessity of its application in a combined-modality. Herein, we aimed to investigate whether c-Myc inhibition could reinforce the anti-leukemic effect of ATO, while reducing its concentration in APL cells. Methods: NB4 cells were treated with the relevant concentrations of 10058-F4 (c-Myc inhibitor) and ATO, and then the survival of the cells was evaluated using trypan blue, MTT and BrdU assays. Moreover, the mechanism of action of the agents were evaluated using Flow cytometry, qRT-PCR and western blot analysis. Results: We found that the inhibition of c-Myc using 10058-F4 could enhance the anti-leukemic effect of ATO in APL cells through reducing the phosphorylation of IκB, decreasing the expression of the anti-apoptotic genes and in turn, inducing a caspase-3-dependent apoptotic cell death. Moreover, the combination of 10058-F4 and ATO abrogated the activation of the PI3K pathway, while neither agent had significant suppressive impact on this pathway; suggesting for the first time that probably the companionship of c-Myc inhibitor may be an appealing strategy for shifting the resistance condition toward a chemo-sensitive phenotype, without the necessity to elevate the effective dose of ATO. Conclusion: Given the efficacy of 10058-F4 in adjuvanting approaches, we suggest this small molecule inhibitor as an impressing agent to be used alongside ATO in the treatment of APL. © 2020 IMS

Investigation of correlation between H63D and C282Y mutations in HFE gene and serum Ferritin level in beta-thalassemia major patients �tude de la corrélation entre les mutations H63D et C282Y du gène HFE et du taux sérique de ferritine chez des patients bêta-thalassémiques majeurs

Introduction: Mutations in the HFE gene have been shown to be associated with hemochromatosis which is observed in beta-thalassemia major. In this study, we determined the HFE gene mutations (C282Y and H63D) among b-thalassemia major patients to investigate the effect of these mutations on serum Ferritin levels. Material and methods: In this cross-sectional study, a total of 105 b-thalassemia subjects with a history of regular blood transfusion were selected. They divided into two distinct groups according cut off 1000 ng/ml of serum Ferritin levels. The HFE gene mutant allele detected by RFLP-PCR. Results: Of 105 thalassemia patients, 29 patients (14 male and 15 female) were heterozygote for H63D mutation, and just one male was homozygote, but for C282Y mutation just one heterozygote and one homozygote was detected, and overall 31 had coexistence of b-thal and HFE gene mutations. As expected, Ferritin levels significantly differed between groups (P = 0.001). Conclusion: The impact of detection of HFE mutations could prognosis the likelihood of iron overload in multi-transfused patients, and allowing early diagnosis and proper management to overcome complications of iron overload in beta-thalassemia patients. © 201

Antileukemic effects of neurokinin-1 receptor inhibition on hematologic malignant cells: A novel therapeutic potential for aprepitant

Genetic and laboratory studies have remodeled the conventional understanding of cancer pathogenesis by identifying different molecular alterations. Intrigued by the contribution of neurokinin-1 receptor (NK1R) network in cancer pathogenesis, we investigated the antileukemic effects of aprepitant, a nonpeptide antagonist of NK1R, in a panel of hematological cell lines. In this study, we found that aprepitant decreased the survival of all the tested cells; however, as compared with NB4, viability of the other cell lines was inhibited at higher concentrations. By increasing both p21 and p73 along with suppressing c-Myc and hTERT, aprepitant probably disordered cell distribution in the cell cycle, decreased DNA replication rate, and, thereby, impeded the proliferative capability of NB4 cells. Moreover, exposing cells to this agent led to activation of the caspase-3-dependent apoptotic pathway through altering the expression of apoptosis-related genes. Noteworthy, aprepitant also sensitized NB4 cells to the cytotoxic effects of arsenic trioxide and vincristine. Overall, it seems that pharmaceutical targeting of NK1R using aprepitant, either as a single agent or in combination, possesses novel promising potential for leukemia treatment strategies. Copyright © 2018 Wolters Kluwer Health, Inc

Antileukemic effects of neurokinin-1 receptor inhibition on hematologic malignant cells: A novel therapeutic potential for aprepitant

Genetic and laboratory studies have remodeled the conventional understanding of cancer pathogenesis by identifying different molecular alterations. Intrigued by the contribution of neurokinin-1 receptor (NK1R) network in cancer pathogenesis, we investigated the antileukemic effects of aprepitant, a nonpeptide antagonist of NK1R, in a panel of hematological cell lines. In this study, we found that aprepitant decreased the survival of all the tested cells; however, as compared with NB4, viability of the other cell lines was inhibited at higher concentrations. By increasing both p21 and p73 along with suppressing c-Myc and hTERT, aprepitant probably disordered cell distribution in the cell cycle, decreased DNA replication rate, and, thereby, impeded the proliferative capability of NB4 cells. Moreover, exposing cells to this agent led to activation of the caspase-3-dependent apoptotic pathway through altering the expression of apoptosis-related genes. Noteworthy, aprepitant also sensitized NB4 cells to the cytotoxic effects of arsenic trioxide and vincristine. Overall, it seems that pharmaceutical targeting of NK1R using aprepitant, either as a single agent or in combination, possesses novel promising potential for leukemia treatment strategies. Copyright © 2018 Wolters Kluwer Health, Inc

Apoptotic Effect of Phosphatidylinositol 3-Kinase Inhibition on Acute Lymphoblastic Leukemia Cells Using Buparlisib

BACKGROUND AND OBJECTIVE: Resistance to chemotherapy is one of the most important problems in treatment of patients diagnosed with acute lymphoblastic leukemia (ALL). Pathway interruption of the phosphatidylinositol -3 kinase (PI3K) and its relation to resistance phenomena cause the inhibitors of this pathway, particularly buparlisib are introduced as one of the most promising cancer drugs. The aim of this study was to evaluate the effect of PI3K pathway inhibition on reducing the survival and induction of apoptosis in Nalm-6 cells using buparlisib. METHODS: In this experimental study, the phosphorylation level of Akt was evaluated using western blot to measure the effect of buparlisib on PI3K/Akt pathway in Nalm-6 cells. Nalm-6 cells were treated with different concentrations of buparlisib (0.5-4 &micro;M) for 24, 36 and 48 hours to study the cytotoxic effect of this inhibitor and then, the metabolic activity, induction of apoptosis and changes&nbsp;in&nbsp;expression of genes involved&nbsp;in&nbsp;apoptosis were evaluated using MTT assay, Annexin/PI staining and Rq-PCR, respectively. FINDINGS: Results showed that PI3K pathway inhibition using buparlisib causes the cytotoxic effect on Nalm-6 cells in a dose- and time-dependent manner through reducing p-Akt. These findings suggested that probably, the anti-leukemic effect of buparlisib is mediated through almost 17-fold increase in apoptotic cells (p&le;0.001) and rising the mRNA expression level of pro-apoptotic genes (p&le;0.01). CONCLUSION: The results indicated that buparlisib has anti-tumor activity against Nalm-6 cells so this inhibitor can be used as a promising agent for the treatment of ALL

Suppression of c-Myc using 10058-F4 exerts caspase-3-dependent apoptosis and intensifies the antileukemic effect of vincristine in pre-B acute lymphoblastic leukemia cells

Despite an old history behind the identification of the leading role of c-Myc in leukemogenesis, the road to constructing a therapeutic perspective for this molecule in acute lymphoblastic leukemia (ALL) is yet mesmerizing. This study was designed to provide a better outlook for the anticancer property of 10058-F4, an appealing inhibitor of c-Myc, in pre-B ALL cell lines either in the context of monotherapy or in combination with chemotherapeutic drugs. Our results declared that abrogation of c-Myc decreased the proliferative capacity of pre-B ALL-derived cells through halting the transition of the cells from G1 phase, and reducing the replicative potential of both REH and Nalm-6 cells, at least partly, through c-Myc-mediated suppression of human telomerase reverse transcriptase. Moreover, 10058-F4 potently induced a caspase-3-dependent apoptosis in pre-B ALL cells via shifting the balance between pro- and anti-apoptotic target genes. Although the inhibition of PI3Kδ using Idelalisib upregulated the messenger RNA expression of autophagy-related genes in 10058-F4-treated cells, treatment with autophagy inhibitor chloroquine decreased viability of the cells, either as a single agent or in combination with Idelalisib and/or 10058-F4; suggesting that the activation of autophagy in pre-B ALL cells could blunt apoptotic events and attenuate anticancer effect of both c-Myc and PI3K inhibitors. Finally, the results of our synergistic experiments delineated that 10058-F4 produced a synergistic effect with vincristine and provided an enhanced therapeutic efficacy in ALL cells, highlighting that c-Myc oncoprotein could be a bona fide target for the treatment of ALL. © 2019 Wiley Periodicals, Inc