Neuroblastoma (NB) is the most common childhood cancer arising from the nervous system. Many high-risk neuroblastoma (HRNB) patients develop relapse after initial response to induction treatment and overall long term ...Neuroblastoma (NB) is the most common childhood cancer arising from the nervous system. Many high-risk neuroblastoma (HRNB) patients develop relapse after initial response to induction treatment and overall long term survival remains poor (less than 60%), emphasizing the need for new therapeutic approaches and more effective treatments. Combination therapies present a favorable approach to improve efficacy, decrease toxicity, and reduce development of drug resistance. Difluoromethylornithine (DFMO) has shown promise in recent clinical trials as a therapeutic agent in treating HRNB. Proteasomes are known to play an important role in tumor cell growth. Bortezomib was the first proteasome inhibitor shown to have anticancer activity clinically. In this study we explore the mechanistic and therapeutic effects of the novel drug combination of DFMO and bortezomib in NB. Cell proliferation studies demonstrated synergistic inhibition of NB cell growth. Bortezomib induced cleaved caspase-3 apoptotic pathway whereas DFMO induced a cytostatic effect on NB cells. Western blot analyses demonstrated down regulation of MYCN, LIN28 and NF-kB in response to DFMO and bortezomib, pathways that are important in cancer stem cells. A decrease in ATP-per-cell when treated with combination therapy suggests inhibition of glycolytic metabolism in NB cells. DFMO as a single agent or in combination with bortezomib significantly reduced tumor growth in xenograft mice. Given the lack of effective treatments, DFMO coupled with bortezomib offers a potential new therapeutic treatment for children with NB.展开更多
Objective:This study aimed to elucidate the influence of IFN-gamma(IFN-γ)in neuroblastoma(NB)cells and reveal its potential underlying molecular mechanism.Methods:The Cell Counting Kit-8,Transwell apparatus,and flow ...Objective:This study aimed to elucidate the influence of IFN-gamma(IFN-γ)in neuroblastoma(NB)cells and reveal its potential underlying molecular mechanism.Methods:The Cell Counting Kit-8,Transwell apparatus,and flow cytometry were employed to assess cellular viability,migratory capacity,invasive potential,and apoptotic rates,respectively.RNA-seq combined with bioinformatics analysis revealed differentially expressed genes(DEGs)and their possible biological functions.Protein levels were determined by western blot analysis.Results:IFN-γtreatment resulted in diminished cell viability,mitigated migratory and invasive capabilities,and augmented apoptotic activity in the SK-N-BE(2)cell line,whereas it exhibited the opposite effect in SH-SY5Y cells.Furthermore,interferon regulatory factor 1(IRF-1)was the common DEG in both IFN-γ-treated SK-N-BE(2)and SH-SY5Y cells.Additionally,we found that it was underexpressed in NB tissues.The depletion of IRF-1 promoted the progression of both SK-N-BE(2)and SH-SY5Y cells.Moreover,IRF-1 knockdown effectively counteracted the effects of IFN-γon SK-N-BE(2)cells,while exacerbating them in SH-SY5Y cells.Conclusion:This study verified that IFN-γexerted a distinct role in both N-Myc-and non-N-Myc-amplified NB cells,partially by mediating the expression of IRF-1,suggesting that it may serve as a potent agent for treating patients with NB.展开更多
文摘Neuroblastoma (NB) is the most common childhood cancer arising from the nervous system. Many high-risk neuroblastoma (HRNB) patients develop relapse after initial response to induction treatment and overall long term survival remains poor (less than 60%), emphasizing the need for new therapeutic approaches and more effective treatments. Combination therapies present a favorable approach to improve efficacy, decrease toxicity, and reduce development of drug resistance. Difluoromethylornithine (DFMO) has shown promise in recent clinical trials as a therapeutic agent in treating HRNB. Proteasomes are known to play an important role in tumor cell growth. Bortezomib was the first proteasome inhibitor shown to have anticancer activity clinically. In this study we explore the mechanistic and therapeutic effects of the novel drug combination of DFMO and bortezomib in NB. Cell proliferation studies demonstrated synergistic inhibition of NB cell growth. Bortezomib induced cleaved caspase-3 apoptotic pathway whereas DFMO induced a cytostatic effect on NB cells. Western blot analyses demonstrated down regulation of MYCN, LIN28 and NF-kB in response to DFMO and bortezomib, pathways that are important in cancer stem cells. A decrease in ATP-per-cell when treated with combination therapy suggests inhibition of glycolytic metabolism in NB cells. DFMO as a single agent or in combination with bortezomib significantly reduced tumor growth in xenograft mice. Given the lack of effective treatments, DFMO coupled with bortezomib offers a potential new therapeutic treatment for children with NB.
基金funded by the Special Basic Cooperative Research Programs of Yunnan Provincial Undergraduate Universities’Association(No.202101BA070001-126).
文摘Objective:This study aimed to elucidate the influence of IFN-gamma(IFN-γ)in neuroblastoma(NB)cells and reveal its potential underlying molecular mechanism.Methods:The Cell Counting Kit-8,Transwell apparatus,and flow cytometry were employed to assess cellular viability,migratory capacity,invasive potential,and apoptotic rates,respectively.RNA-seq combined with bioinformatics analysis revealed differentially expressed genes(DEGs)and their possible biological functions.Protein levels were determined by western blot analysis.Results:IFN-γtreatment resulted in diminished cell viability,mitigated migratory and invasive capabilities,and augmented apoptotic activity in the SK-N-BE(2)cell line,whereas it exhibited the opposite effect in SH-SY5Y cells.Furthermore,interferon regulatory factor 1(IRF-1)was the common DEG in both IFN-γ-treated SK-N-BE(2)and SH-SY5Y cells.Additionally,we found that it was underexpressed in NB tissues.The depletion of IRF-1 promoted the progression of both SK-N-BE(2)and SH-SY5Y cells.Moreover,IRF-1 knockdown effectively counteracted the effects of IFN-γon SK-N-BE(2)cells,while exacerbating them in SH-SY5Y cells.Conclusion:This study verified that IFN-γexerted a distinct role in both N-Myc-and non-N-Myc-amplified NB cells,partially by mediating the expression of IRF-1,suggesting that it may serve as a potent agent for treating patients with NB.
