Childhood neuroblastoma,a leading cause of cancer-related mortality in young children,accounts for approximately 8%-10%of pediatric cancers1.Originating from neural crest cells of the sympathetic nervous system,these ...Childhood neuroblastoma,a leading cause of cancer-related mortality in young children,accounts for approximately 8%-10%of pediatric cancers1.Originating from neural crest cells of the sympathetic nervous system,these tumors affect primarily children younger than 5 years of age and are often diagnosed in advanced stages,because of their aggressive nature and vague early symptoms2-4.展开更多
The published article titled“Triptolide inhibits proliferation and migration of human neuroblastoma SH-SY5Y cells by upregulating microRNA-181a”has been retracted from Oncology Research,Vol.26,No.8,2018,PP.1235-1243.
Objective:Neuroblastoma is the most common extracranial solid tumor in children and has complex genetic underpinnings.Previous genome-wide association studies(GWASs)have identified many loci associated with neuroblast...Objective:Neuroblastoma is the most common extracranial solid tumor in children and has complex genetic underpinnings.Previous genome-wide association studies(GWASs)have identified many loci associated with neuroblastoma susceptibility;however,their application in risk prediction for Chinese children has not been systematically explored.This study seeks to enhance neuroblastoma risk prediction by validating these loci and evaluating their performance in polygenic risk models.Methods:We validated 35 GWAS-identified neuroblastoma susceptibility loci in a cohort of Chinese children,consisting of 402 neuroblastoma patients and 473 healthy controls.Genotyping these polymorphisms was conducted via the TaqMan method.Univariable and multivariable logistic regression analyses revealed the genetic loci significantly associated with neuroblastoma risk.We constructed polygenic risk models by combining these loci and assessed their predictive performance via area under the curve(AUC)analysis.We also established a polygenic risk scoring(PRS)model for risk prediction by adopting the PLINK method.Results:Fourteen loci,including ten protective polymorphisms from CASC15,BARD1,LMO1,HSD17B12,and HACE1,and four risk variants from BARD1,RSRC1,CPZ and MMP20 were significantly associated with neuroblastoma risk.Compared with single-gene model,the 8-gene model(AUC=0.72)and 13-gene model(AUC=0.73)demonstrated superior predictive performance.Additionally,a PRS incorporating six significant loci achieved an AUC of 0.66,effectively stratifying individuals into distinct risk categories regarding neuroblastoma susceptibility.A higher PRS was significantly associated with advanced International Neuroblastoma Staging System(INSS)stages,suggesting its potential for clinical risk stratification.Conclusions:Our findings validate multiple loci as neuroblastoma risk factors in Chinese children and demonstrate the utility of polygenic risk models,particularly the PRS,in improving risk prediction.These results suggest that integrating multiple genetic variants into a PRS can enhance neuroblastoma risk stratification and potentially improve early diagnosis by guiding targeted screening programs for high-risk children.展开更多
Objective:Neuroblastoma(NB)is frequently associated with high-risk pediatric cases that demonstrate limited response to cisplatin,contributing to a poor prognosis.Recent studies have explored the role of tumor cell se...Objective:Neuroblastoma(NB)is frequently associated with high-risk pediatric cases that demonstrate limited response to cisplatin,contributing to a poor prognosis.Recent studies have explored the role of tumor cell senescence in increasing sensitivity to this chemotherapy agent.This study aims to identify genes related to cell senescence in children diagnosed with NB,evaluate their influence on cisplatin sensitivity,and investigate potential strategies to enhance the efficacy of chemotherapy.Methods:Gene expression profiles and clinical data were obtained for 498 NB patients from the GEO database(GSE49710).The study focused on identifying genes that were differentially expressed between stage IV and other stages,particularly those linked to cell senescence and cisplatin resistance.To analyze the prognostic significance of these differentially expressed genes,we employed LASSO regression and multivariate Cox proportional hazards models.Transcriptomic and proteomic analyses of 15 NB specimens revealed a significant correlation between Flap endonuclease-1(FEN1)expression levels and both cellular senescence and sensitivity to cisplatin.We quantified FEN1 expression and cisplatin IC50 values in four different NB cell lines.The influence of FEN1 knockdown and overexpression on NB cell proliferation,invasion,and migration was evaluated using cloning assays,transwell assays,and scratch assays.Furthermore,we utilized Western blotting to analyze senescenceassociated proteins p21 and proliferating cell nuclear antigen(PCNA),thereby evaluating the role of FEN1 in cellular senescence.The impact of FEN1 on cisplatin sensitivity was investigated via the CCK-8 cell counting assay.Additionally,we investigated how FEN1 inhibitors might impact NB cell proliferation and enhance the therapeutic efficacy of cisplatin treatment.Results:FEN1 was found to be highly expressed in stage IV NB and showed a strong association with cisplatin sensitivity,establishing it as a critical molecular marker linked to poor patient prognosis.Notably,elevated FEN1 expression correlated with reduced sensitivity to cisplatin,as evidenced by higher IC50 values.In the SH-SY5Y cell line,FEN1 knockdown led to significant reductions in cell proliferation,invasion,and migration,along with an increase inβ-galactosidase staining—indicative of senescence.This knockdown also resulted in elevated levels of the p21 protein and decreased expression of PCNA,concurrently lowering cisplatin IC50 values.Conversely,FEN1 overexpression in the SK-N-SH cell line resulted in enhanced cell proliferation,invasion,and migration.This overexpression was associated with reducedβ-galactosidase staining,decreased levels of p21,and increased expression of PCNA,ultimately resulting in higher cisplatin IC50 values.Importantly,FEN1 inhibitors alone significantly impeded NB cell proliferation,and their combination with cisplatin further amplified this inhibitory effect compared to cisplatin treatment alone.Conclusions:Bioinformatics and sequencing analyses indicate that the senescence-related gene FEN1 is significantly associated with cisplatin sensitivity and adverse prognosis in pediatric NB.FEN1 plays a pivotal role in regulating NB cell proliferation,invasion,and migration,thereby facilitating cancer progression.Furthermore,it influences cisplatin sensitivity through its effects on cellular senescence.FEN1 inhibitors demonstrate potential both as monotherapies and in conjunction with cisplatin,suggesting that targeting FEN1 may be represent a valuable strategy for improving outcomes in high-risk NB patients.展开更多
BACKGROUND Neuroblastoma(NB)is one of the most common malignancies in children.Metastasis in NB is not uncommon.However,nasal metastases are rare.Here,we reported two pediatric cases of nasal metastases.CASE SUMMARY C...BACKGROUND Neuroblastoma(NB)is one of the most common malignancies in children.Metastasis in NB is not uncommon.However,nasal metastases are rare.Here,we reported two pediatric cases of nasal metastases.CASE SUMMARY Case 1 was a 3-year-old boy without a history of NB.Case 2 was a 10-year-old girl who had a history of NB for 6 years.Both of them presented with symptoms of nasal and sinus masses such as epistaxis or discharge from the nose.The radiologic imaging results revealed masses in the nasal cavity or nasopharynx in both cases and a mass in the right adrenal gland of case 1.The pathologic examination of biopsy samples of their nasal masses revealed“small round bluecell tumor”along with abundant vascular fibrous septa.The tumor cells expressed synaptophysin,cluster of differentiation 56,chromogranin A,paired like homeobox protein 2B and a very high Ki67 index in both case but were negative for vimentin,desmin,leucocyte common antigen and cytokeratin.Myelocytomatosis viral related oncogene,neuroblastoma derived(MYCN)amplification was detected in both cases.Finally,the two cases were diagnosed as nasal metastases from NB based on the clinical and pathologic findings.The two patients affected by NB were>18 mo old,the primary tumor location was adrenal gland,and they presented with multiple metastases.CONCLUSION It is difficult to differentiate between metastatic NB in the nose and olfactory neuroblastoma in the absence of a history of NB.Paired like homeobox protein 2B can play an important role in the diagnosis and differential diagnosis of this disease.展开更多
Esthesioneuroblastoma is a malignant tumor, arising in the upper nasal cavity, that could spread to the frontal lobe of the brain as well as metastasize to the lymph nodes. Due to the low incidence of this tumor, FDA-...Esthesioneuroblastoma is a malignant tumor, arising in the upper nasal cavity, that could spread to the frontal lobe of the brain as well as metastasize to the lymph nodes. Due to the low incidence of this tumor, FDA-approved treatment modalities do not exist and clinical trials have not been performed. We present an interesting case of a 66-year-old female, diagnosed with Kadish stage B esthesioneuroblastoma and stage IIA nonsmall cell carcinoma of the lung, who benefited from our treatment. Both malignancies were diagnosed in 2002 at which time the patient consented to undergo left upper lobectomy for her lung cancer, but she refused the craniofacial resection and radiation therapy recommended for treatment of her esthesioneuroblastoma. From 2003 to 2004 she received treatment at the Burzynski Clinic with oral sodium phenylbutyrate (0.2 g/kg/day). She tolerated the treatment very well without significant adverse events. Gradual reduction in her tumor size was confirmed by repeat MRIs. From treatment start in March 2003 to December 2003 her tumor decreased by 40%. Subsequent MRI from March 2004 revealed increased tumor size, which, however, was still a 13% reduction from the baseline MRI. What is important to mention is that in addition to shrinkage of the esthesioneuroblastoma, the patient obtained the clinical benefit of 3.5-years longer survival than was predicted for her lung cancer—whereas the median survival for a patient with stage IIA adenocarcinoma of the left upper lobe of the lung is approximately two years, our patient survived more than five and a half years. The effect of phenylbutyrate (PB) and its metabolite phenylacetate on neuroblastoma and lung cancer is documented by numerous preclinical studies and is also evident in this case. It is proposed that the activity of these two compounds is mediated through increased expression of the p21 tumor suppressor gene. p21 is a strong inhibitor of cyclin-D and cyclin-dependent kinase 4, which contribute to undifferentiated phenotype in neuroblastoma and are instrumental in cell cycle progression from G1 to S phase. It is hoped that future research and combination of PB with other chemotherapeutic and targeted agents will provide better control of esthesioneuroblastoma and lung cancer.展开更多
Olfactory neuroblastoma (esthesioneuroblastoma, ЕNB) is a rare tumor arising from the olfactory neuroepithelium. We report a case of ЕNB located in inferior nasal concha, combined with thyroid gland carcinoma and ga...Olfactory neuroblastoma (esthesioneuroblastoma, ЕNB) is a rare tumor arising from the olfactory neuroepithelium. We report a case of ЕNB located in inferior nasal concha, combined with thyroid gland carcinoma and gastrointestinal stromal carcinoma in a 77-year-old man. The tumor was resected endonasally. When the final diagnosis of olfactory neuroblastoma was confirmed by histopathologic examination and immunohistochemical staining, the PET/CT examination was performed. The imaging revealed a small focus of a moderately increased cancer activity in the thyroid region. A gastrointestinal stromal carcinoma was detected one year after the resection of the thyroid gland. We discuss the clinical appearance of ENB, staging systems, diagnosis and management. During the endonasal surgery, ENB was removed entirely. Seven days after operation, in order to monitor the postoperative result, PET/CT was performed and a papillary thyroid cancer was detected. One year after the thyroid surgery, gastroendoscopy showed a neoplastic formation in the stomach. In conclusion, we state that when identified as aggressive tumors such as ENB, it is necessary to provide regular examinations in order to detect distant ENB metastases or other neoplastic localisations.展开更多
This article presents a case of a patient with relapsed esthesioneuroblastoma (ENB), an aggressive rare tumor that arises from the specialized sensory epithelial olfactory cells in the skull base area, which was initi...This article presents a case of a patient with relapsed esthesioneuroblastoma (ENB), an aggressive rare tumor that arises from the specialized sensory epithelial olfactory cells in the skull base area, which was initially treated with endoscopic surgery, followed by adjuvant radiotherapy. After local relapse, new surgical approaches and subsequent lines of platin-based chemotherapy were performed. A PET-CT with <sup>68</sup>GALIUM DOTATATOC (PET-DOTATOC) showed intense uptake of disease, compatible with the presence of somatostatin receptors, in the face, nodes, liver, bones, and meningeal area. Treatment with 4 cycles of <sup>177</sup>Lutetium-Dotatate was performed, followed by maintenance octreotide, with a major radiological and clinical response that is lasting more than 1 year after treatment. This article describes a rare case of a skull-base tumor, with multiple recurrences, in which disease control was achieved with a targeted Peptide Receptor Radionuclide Therapy (PRRT) with <sup>177</sup>Lutetium-Dotatate, and discusses factors that could influence the incorporation of this form of therapy. Previous case reports proved the potential efficacy of this therapy usually given for low-grade neuroendocrine tumors and will be carefully reviewed.展开更多
Objective:To evaluate the efficacy of allicin combined with cyclophosphamide on neuroblastoma(NB)-bearing mice and explore the immunological mechanism in it.Methods:A total of 30NB-bearing mice were equally randomized...Objective:To evaluate the efficacy of allicin combined with cyclophosphamide on neuroblastoma(NB)-bearing mice and explore the immunological mechanism in it.Methods:A total of 30NB-bearing mice were equally randomized into model group,cyclophosphamide group and combined therapy group,10 nudemice were set as normal saline(NS) group.Cyclophosphamide group and combined therapy group were weekly injected with 60 mg/kg cyclophosphamide for four weeks;besides,combined therapy group was given with allicin(10 mg/kg/d) by gastric perfusion for 4 weeks;model group and NS group were given with the same volume of NS.Serum VEGF content was detected by ELISA pre-treating(0 d) and on the 3rd d.14 th d and 28 th d;on29th d,all mice were sacrificed and the tumor,liver,spleen and thymic tissues were weighted.Tumors were made into paraffin section for detecting tumor cell apoptosis and proliferation by TUNEI.and BrdU method,respectively.Survival curves were drawn by Kaplan-Meier method.Results:After treatment,both treatment groups relieved on viscera indexes,VEGF level,T cell subsets distribution and tumor growth and each index of combined therapy group was better than cyclophosphamide group(P【0.05 or 0.01);only combined therapy group could significantly increase the lifetime of NB-bearing mice(χ~2=5.667,P=0.017).Conclusions:Allicin can improve T cell subsets distribution and inhibit VEGF expression through its immunomodulatory activity,thereby improve the efficiency on NB in coordination with cyclophosphamide.展开更多
Neuroblastoma(NB) is the most common extracranial solid tumor in children. Diarrheal NB is quite rare and is not easy to diagnose in the early stage. Six cases of diarrheal NB in our hospital treated from 1996 to 2006...Neuroblastoma(NB) is the most common extracranial solid tumor in children. Diarrheal NB is quite rare and is not easy to diagnose in the early stage. Six cases of diarrheal NB in our hospital treated from 1996 to 2006 were retrospectively analyzed, including characteristics such as electrolyte imbalance, pathologic features, vasoactive intestinal peptide(VIP) immunohistochemical staining results, treatment, and prognosis. All patients were boys with 3-8 loose or watery stools each day and routine fecal tests were normal. Abdominal tumors were identified by B-ultrasound. Drugs were ineffective. Three patients underwent surgery, and the remaining three patients received surgery and chemotherapy. Diarrhea stopped after treatment in five patients. Two patients died due to intractable hypokalemia. The tumor was located in the adrenal gland in four patients, in the upper retroperitoneum in one patient, and in the presacral area in one patient. Pathologic findings were NB and ganglioneuroblastoma. Five patients were at clinical stage Ⅰ-Ⅱ, and one was at stage Ⅲ. Four patients survived(followed-up for 6 mo to 4 years). Immunohistochemical staining for VIP was positive. Refractory diarrhea is a paraneoplastic syndrome of NB and is rare. Patients aged 1-3 years who present with chronic intractable diarrhea should be followed closely. Intractable diarrhea, hypokalemia, and dysplasia are the initial clinical manifestations. Increased VIP is characteristic of this disease. Potassium supplementation plays a vital role in the treatment procedure, especially preoperatively. The prognosis of diarrheal NB is good following appropriate treatment.展开更多
Introduction: Brain metastasis is common in relapsed neuroblastoma patients, but the characteristics of brain metastasis remain largely unknown. This study aimed to investigate the status of brain metastasis with neur...Introduction: Brain metastasis is common in relapsed neuroblastoma patients, but the characteristics of brain metastasis remain largely unknown. This study aimed to investigate the status of brain metastasis with neuroblastoma in South China.Methods: In this retrospective case?based study, 106 patients with stage 4 neuroblastoma from the Department of Pediatric Oncology in Sun Yat?sen University Cancer Center between January 2004 and May 2013 were included. The incidence, risk factors, and survival status of these patients were reviewed and analyzed.Results: Of the 106 patients, 11(10.4%) developed brain metastasis, accounting for 20.0% of 55 patients with relapse or progression. The age at initial diagnosis of the 11 patients ranged from 2 to 10 years(median 4 years), which was younger than that of the patients without brain metastasis(median 5 years, range 1–10 years, P = 0.073). The male to female ratio of the 11 patients was 8:3, which was not signiicantly diferent from that of the patients with?out brain metastasis(P = 0.86). Patients with brain metastasis had higher lactate dehydrogenase levels than those without brain metastasis, but the diferences were not signiicant(P initial diagnosis to the develo= 0.076). Eight patients died, and 3 patients survived. The median interval from thepment of brain metastasis was 18 months(range 6–32 months). The median survival was 4 months(range 1 day to 29 months) after the diagnosis of brain metastasis. The median interval from the manifestation of brain metastasis to death was 3 months(range 1 day to 11 months).Conclusions: High?risk factors for brain metastasis in cases of neuroblastoma include bone marrow involvement and a younger age at initial diagnosis. Nevertheless, multiple treatment modalities can improve disease?free survival.展开更多
Thymoquinone(TQ), an active component derived from the medial plant Nigella sativa, has been used for medical purposes for more than 2 000 years. Recent studies have reported that TQ blocked angiogenesis in animal mod...Thymoquinone(TQ), an active component derived from the medial plant Nigella sativa, has been used for medical purposes for more than 2 000 years. Recent studies have reported that TQ blocked angiogenesis in animal model and reduced migration, adhesion, and invasion of glioblastoma cells. We have recently shown that TQ could exhibit a potent cytotoxic effect and induce apoptosis in mouse neuroblastoma(Neuro-2a) cells. In the present study, TQ treatment markedly decreased the adhesion and migration of Neuro-2a cells. TQ down-regulated MMP-2 and MMP-9 protein expression and m RNA levels and their activities. Furthermore, TQ significantly down-regulated the protein expression of transcription factor NF-κB(p65) but not significantly altered the expression of N-Myc. Taken together, our data indicated that TQ's inhibitory effect on the migration of Neuro-2a cells was mediated through the suppression of MMP-2 and MMP-9 expression, suggesting that TQ treatment can be a promising therapeutic strategy for human malignant neuroblastoma.展开更多
This study investigated the effects of small interfering RNA （siRNA）-mediated silencing of chemokine receptor 4 （CXCR4） on the invasion capacity of human neuroblastoma cell line SH-SY5Y in vitro. Three siRNAs targ...This study investigated the effects of small interfering RNA （siRNA）-mediated silencing of chemokine receptor 4 （CXCR4） on the invasion capacity of human neuroblastoma cell line SH-SY5Y in vitro. Three siRNAs targeting CXCR4 were chemically synthesized and individually transfected into SH-SY5Y cells. Expression of CXCR4 mRNA and protein was signiifcantly sup-pressed in transfected cells by all three sequence-speciifc siRNAs compared with control groups. Furthermore, the invasion capacity of SH-SY5Y cells was signiifcantly decreased following trans-fection with CXCR4-speciifc siRNA compared with the control groups. These data demonstrate that down-regulation of CXCR4 can inhibit in vitro invasion of neuroblastoma.展开更多
Objective To study the effect of γ-interferon （IFNγ）, tumor necrosis factor related apoptosis inducing ligand （TRAIL）, and cisplatin or etoposide induced apoptosis in human neuroblastoma cell line SH-SY5Y and it...Objective To study the effect of γ-interferon （IFNγ）, tumor necrosis factor related apoptosis inducing ligand （TRAIL）, and cisplatin or etoposide induced apoptosis in human neuroblastoma cell line SH-SY5Y and its possible molecular mechanisms. Methods The expressions of Caspase 8 mRNA and protein were detected with RT-PCR and Western blot analysis. The effects of IFNγ, TRAIL, IFNγ ＋ TRAIL, IFNγ ＋ Caspase 8 inhibitor ＋ TRAIL, IFNγ ＋ cisplatin ＋ TRAIL, and IFNγ ＋ etoposide ＋ TRAIL on the growth and apoptosis of SH-SY5Y cells were detected with the methods of MTT and flow cytometry. The relative Caspase 8 activity was measured with colorimetric assay. Results Caspase 8 was undetectable in SH-SY5Y cells but an increased expression of Caspase 8 mRNA and protein was found after treatment with IFNγ. SH-SY5Y ceils themselves were not sensitive to TRAIL, but those expressing Caspase 8 after treatment with IFNγ were. The killing effect of TRAIL on SH-SY5Y cells expressing Caspase 8 was depressed by Caspase 8 inhibitor. Cisplatin and etoposide could enhance the sensitivity of TRAIL on SH-SY5Y cells. The relative Caspase 8 activity of SH-SY5Y cells in IFNγ ＋ TRAIL group was significantly higher than those of control group, IFNγ group, TRAIL group, and inhibitor group （ P 〈 0. 01 ）. There was no significant difference among IFNγ ＋ TRAIL group, IFNγ ＋ cisplatin ＋ TRAIL group, and IFNγ ＋ etoposide ＋ TRAIL group. Conclusions IFNγ could sensitize SH-SY5Y cells to TRAIL-induced apoptosis and this may be realized by the up-regulation of Caspase 8. Cisplatin and etoposide could enhance the killing effect of TRAIL on SH-SY5Y cells.展开更多
Objective:AlkB homolog 5(ALKBH5)has been proven to be closely related to tumors.However,the role and molecular mechanism of ALKBH5 in neuroblastomas have rarely been reported.Methods:The potential functional single-nu...Objective:AlkB homolog 5(ALKBH5)has been proven to be closely related to tumors.However,the role and molecular mechanism of ALKBH5 in neuroblastomas have rarely been reported.Methods:The potential functional single-nucleotide polymorphisms(SNPs)in ALKBH5 were identified by National Center for Biotechnology Information(NCBI)dbSNP screening and SNPinfo software.TaqMan probes were used for genotyping.A multiple logistic regression model was used to evaluate the effects of different SNP loci on the risk of neuroblastoma.The expression of ALKBH5 in neuroblastoma was evaluated by Western blotting and immunohistochemistry(IHC).Cell counting kit-8(CCK-8),plate colony formation and 5-ethynyl-2'-deoxyuridine(EdU)incorporation assays were used to evaluate cell proliferation.Wound healing and Transwell assays were used to compare cell migration and invasion.Thermodynamic modelling was performed to predict the ability of miRNAs to bind to ALKBH5 with the rs8400 G/A polymorphism.RNA sequencing,N6-methyladenosine(mA)sequencing,mA methylated RNA immunoprecipitation(MeRIP)and a luciferase assay were used to identify the targeting effect of ALKBH5 on SPP1.Results:ALKBH5 was highly expressed in neuroblastoma.Knocking down ALKBH5 inhibited the proliferation,migration and invasion of cancer cells.miR-186-3p negatively regulates the expression of ALKBH5,and this ability is affected by the rs8400 polymorphism.When the G nucleotide was mutated to A,the ability of miR-186-3p to bind to the 3'-UTR of ALKBH5 decreased,resulting in upregulation of ALKBH5.SPPI is the downstream target gene of the ALKBH5 oncogene.Knocking down SPP1 partially restored the inhibitory effect of ALKBH5 downregulation on neuroblastoma.Downregulation of ALKBH5 can improve the therapeutic efficacy of carboplatin and etoposide in neuroblastoma.Conclusions:We first found that the rs8400 G>A polymorphism in the m6A demethylase-encoding gene ALKBH5 increases neuroblastoma susceptibility and determines the related mechanisms.The aberrant regulation of ALKBH5 by miR-186-3p caused by this genetic variation in ALKBH5 promotes the occurrence and development of neuroblastoma through the ALKBH5-SPP1 axis.展开更多
Objective： Id2 is a natural inhibitor of the basic helix-loop-helix（bHLH） transcription factors. Although it is well known that active ld2 prevents differentiation and promotes cell cycle progression and tumorigene...Objective： Id2 is a natural inhibitor of the basic helix-loop-helix（bHLH） transcription factors. Although it is well known that active ld2 prevents differentiation and promotes cell cycle progression and tumorigenesis, the molecular events that regulate Id2 activity remain to be investigated. Methods： Yeast two-hybrid, mammalian two-hybrid, GST-pulldown and immunoprecipitation （ColP） assays were used to screen and identify novel Id2 interactors. Luciferase assays were used to detect E47-mediated transcription activity. Colony formation and BrdU incorporation assays were used to determine cellular proliferation abilities. Northorn blot, western blot and quantitative PCR methods were used to measure gene expression levels. Electrophoretic mobility shift assays （EMSAs） were performed to investigate protein/DNA binding. Results： The LIM-only protein FHL2 （four-and-a-half-LIM-only protein 2） was identified to be a novel Id2 interactor. The HLH domain within Id2 is not required for its interaction with FHL2. FHL2 antagonizes the inhibitory effect of Id2 on the basic helix-loop-helix protein E47-mediated transcription. FHL2 prevents the formation of Id2-E47 heterdimer, thus releasing E47 to its target DNA and restoring its transcriptional activity. FHL2 expression was remarkably up-regulated during retinoic acid-induced differentiation of neuroblastoma cells, during which the expression of Id2 is opposite to that. Ectopic FHL2 expression in neuroblastoma cells markedly reduces the transcriptional and cell-cycle promoting functions of Id2. Conclusion： These results indicate that FHL2 is an important repressor of the oncogenic activity of Id2 in neuroblastoma cells.展开更多
The active ingredient of ginseng,ginsenosides Rg1,has been shown to scavenge free radicals and improve antioxidant capacity.This study hypothesized that ginsenosides Rg1 has a protective role in human neuroblastoma ce...The active ingredient of ginseng,ginsenosides Rg1,has been shown to scavenge free radicals and improve antioxidant capacity.This study hypothesized that ginsenosides Rg1 has a protective role in human neuroblastoma cells injured by H2O2.Ginsenosides Rg1 at different concentrations（50 and 100 μM） was used to treat H2O2（150 μM）-injured SH-SY5 Y cells.Results demonstrated that ginsenoside Rg1 elevated the survival rate of SH-SY5 Y cells injured by H2O2,diminished the amount of leaked lactate dehydrogenase,and increased superoxide dismutase activity.Ginsenoside Rg1 effectively suppressed caspase-3 immunoreactivity,and contributed to heat shock protein 70 gene expression,in a dose-dependent manner.These results indicate that ginsenoside Rg1 has protective effects on SH-SY5 Y cells injured by H2O2 and that its mechanism of action is associated with anti-oxidation and the inhibition of apoptosis.展开更多
Most therapeutic protocols for child cancers use cytotoxic agents which have a narrow therapeutic index,and resulting in severe acute and chronic toxicities to normal tissues. Despite the fact that most child cancer p...Most therapeutic protocols for child cancers use cytotoxic agents which have a narrow therapeutic index,and resulting in severe acute and chronic toxicities to normal tissues. Despite the fact that most child cancer patients achieve complete remission after chemotherapy,death still occurs due to relapse of persistent minimal residual disease(MRD) which remaining after initial cytotoxic chemotherapy. Advanced neuroblastoma(NB) is a leading cause of cancer deaths in young children. Retinoids are an important component of advanced NB therapy at the stage of MRD,yet half of all patients treated with 13-cis-retinoic acid still relapse and die. More effective combination therapies,with a lower side-effect profile,are required to improve outcomes for NB. Fenretinide or N-4-hydroxyphenyl retinamide is a synthetic derivative of retinoic acid which works on cancer cells through nuclear receptor-dependent and-independent signalling mechanisms. Moreover,several histone deacetylase inhibitors have entered early phase trials,and,suberoylanilide hydroxamic acid has been approved for use in adult cutaneous T cell lymphoma. A number of studies suggest that retinoid signal activation is necessary for histone deacetylase inhibitor activity. A better understanding of their mechanism of actions will lead to more evidence-based retinoid combination therapies.展开更多
Cancers are characterized by deregulation of multiple signaling pathways and thus monotherapies are hardly effective. Neuroblastoma, which often occurs in adrenal glands, is the most common childhood malignancy. Malig...Cancers are characterized by deregulation of multiple signaling pathways and thus monotherapies are hardly effective. Neuroblastoma, which often occurs in adrenal glands, is the most common childhood malignancy. Malignant neuroblastoma resists traditional treatments and further studies are needed for effective therapeutic interventions. We evaluated synergistic efficacy of N-(4-hydroxyphenyl) retinamide (4-HPR) and genistein (GST) for induction of apoptosis in human malignant neuroblastoma SH-SY5Y and SK-N-BE2 cells in culture and activation of multiple pathways for increasing apoptosis in ectopic and orthotopic neuroblastoma xenografts in nude mice. Combination of 4-HPR and GST synergistically reduced cell viability, caused subG1 accumulation, increased caspase-3 activity for apoptosis in vitro and reduced tumor growth in vivo. Western blotting indicated that combination therapy down regulated Id2 to induce differentiation, increased pro-apoptotic Bax and decreased anti-apoptotic Bcl-2 leading to an increase in Bax:Bcl-2 ratio, increased mitochondrial Bax level, caused mitochondrial release of Smac/Diablo, down regulation of the baculovirus inhibitor-of-apoptosis repeat containing (BIRC) proteins such as BIRC-2 and BIRC-3, and activation of calpain and caspase-3 in SH-SY5Y xenografts. Accumulation of apoptosis-inducing-factor (AIF) in cytosol and increase in caspase-4 activation suggested involvement of mitochondrial pathway and endoplasmic reticulum (ER) stress, respectively, for apoptosis in SH-SY5Y xenografts. In situ immunofluorescent labelings of SH-SY5Y and SK-N-BE2 xenograft sections showed overexpression of calpain, caspase-12, and caspase-3, and AIF, suggesting induction of mitochondrial caspase-dependent and caspase-independent pathways for apoptosis. Collectively, synergistic effects of 4-HPR and GST induced mitochondrial pathways and also ER stress for increasing apoptosis in ectopic and orthotopic neuroblastoma xenografts in nude mice.展开更多
The effects of tissue factor （TF） on doxorubicin-induced apoptosis in human neuroblastoma were investigated. The expression of TF was examined by Western blotting. TFsiRNA-pSUPER plasmid was constructed by inserting...The effects of tissue factor （TF） on doxorubicin-induced apoptosis in human neuroblastoma were investigated. The expression of TF was examined by Western blotting. TFsiRNA-pSUPER plasmid was constructed by inserting specific 19-nt silencing sequence targeting TF gene into pSUPER vector. Transfection of TFsiRNA-pSUPER was performed using lipofectamine^2000. The cytotoxicity of doxorubicin was determined by WST assay. The activation of Caspase-3 and PARP induced by doxorubicin was tested by Western blotting. The apoptotic cells were stained by Hochest33342 and counted under fluorescence inverted microscope. It was found that human neuroblastoma cell line SK-N-MC expressed high level of TE Knockdown of the TF expression was achieved by transfection of TFsiRNA-pSUPER on SK-N-MC cells in a dose-dependent manner. Inhibition of TF significantly decreased the viability of transfected SK-N-MC cells treated with different concentrations of doxorubicin. Cleavage of Caspase-3 and PARP was enhanced in transfected SK-N-MC cells with down-regulation of TF. TFsiRNA treatment significantly increased the number of apoptotic cells in transfected SK-N-MC cells as compared with those control cells （P〈0.05） when these cells were exposed to 1 μg/mL doxorubicin for 8 h. These results suggested that knockdown of the TF expression by specific siRNA vector could increase the cytotoxicity of doxorubicin and enhance doxorubicin-induced apoptosis in human neuroblastoma cells. Over-expression of TF might contribute to chemotherapy resistance in human neuroblastoma and its progression, at lest in part, by regulating doxorubicin-induced apoptosis.展开更多
基金supported by grants from the National Natural Science Foundation of China(Grant No.82470544)the Fundamental Research Funds for the Central Universities(Grant No.226-2024-00153).
文摘Childhood neuroblastoma,a leading cause of cancer-related mortality in young children,accounts for approximately 8%-10%of pediatric cancers1.Originating from neural crest cells of the sympathetic nervous system,these tumors affect primarily children younger than 5 years of age and are often diagnosed in advanced stages,because of their aggressive nature and vague early symptoms2-4.
文摘The published article titled“Triptolide inhibits proliferation and migration of human neuroblastoma SH-SY5Y cells by upregulating microRNA-181a”has been retracted from Oncology Research,Vol.26,No.8,2018,PP.1235-1243.
基金supported by grants from the National Natural Science Foundation of China(No.82173593,32300473)Guangzhou Science and Technology Project(No.2025A04J4537,2025A04J4696)+1 种基金Guangdong Basic and Applied Basic Research Foundation(No.2023A1515220053)Postdoctoral Science Foundation of Jiangsu Province(No.2021K524C).
文摘Objective:Neuroblastoma is the most common extracranial solid tumor in children and has complex genetic underpinnings.Previous genome-wide association studies(GWASs)have identified many loci associated with neuroblastoma susceptibility;however,their application in risk prediction for Chinese children has not been systematically explored.This study seeks to enhance neuroblastoma risk prediction by validating these loci and evaluating their performance in polygenic risk models.Methods:We validated 35 GWAS-identified neuroblastoma susceptibility loci in a cohort of Chinese children,consisting of 402 neuroblastoma patients and 473 healthy controls.Genotyping these polymorphisms was conducted via the TaqMan method.Univariable and multivariable logistic regression analyses revealed the genetic loci significantly associated with neuroblastoma risk.We constructed polygenic risk models by combining these loci and assessed their predictive performance via area under the curve(AUC)analysis.We also established a polygenic risk scoring(PRS)model for risk prediction by adopting the PLINK method.Results:Fourteen loci,including ten protective polymorphisms from CASC15,BARD1,LMO1,HSD17B12,and HACE1,and four risk variants from BARD1,RSRC1,CPZ and MMP20 were significantly associated with neuroblastoma risk.Compared with single-gene model,the 8-gene model(AUC=0.72)and 13-gene model(AUC=0.73)demonstrated superior predictive performance.Additionally,a PRS incorporating six significant loci achieved an AUC of 0.66,effectively stratifying individuals into distinct risk categories regarding neuroblastoma susceptibility.A higher PRS was significantly associated with advanced International Neuroblastoma Staging System(INSS)stages,suggesting its potential for clinical risk stratification.Conclusions:Our findings validate multiple loci as neuroblastoma risk factors in Chinese children and demonstrate the utility of polygenic risk models,particularly the PRS,in improving risk prediction.These results suggest that integrating multiple genetic variants into a PRS can enhance neuroblastoma risk stratification and potentially improve early diagnosis by guiding targeted screening programs for high-risk children.
基金supported by grant from Affiliated Hospital of Nantong University(YJXYY202204)Postgraduate Research&Practice Innovation Program of Jiangsu Province(SJCX24_2063).
文摘Objective:Neuroblastoma(NB)is frequently associated with high-risk pediatric cases that demonstrate limited response to cisplatin,contributing to a poor prognosis.Recent studies have explored the role of tumor cell senescence in increasing sensitivity to this chemotherapy agent.This study aims to identify genes related to cell senescence in children diagnosed with NB,evaluate their influence on cisplatin sensitivity,and investigate potential strategies to enhance the efficacy of chemotherapy.Methods:Gene expression profiles and clinical data were obtained for 498 NB patients from the GEO database(GSE49710).The study focused on identifying genes that were differentially expressed between stage IV and other stages,particularly those linked to cell senescence and cisplatin resistance.To analyze the prognostic significance of these differentially expressed genes,we employed LASSO regression and multivariate Cox proportional hazards models.Transcriptomic and proteomic analyses of 15 NB specimens revealed a significant correlation between Flap endonuclease-1(FEN1)expression levels and both cellular senescence and sensitivity to cisplatin.We quantified FEN1 expression and cisplatin IC50 values in four different NB cell lines.The influence of FEN1 knockdown and overexpression on NB cell proliferation,invasion,and migration was evaluated using cloning assays,transwell assays,and scratch assays.Furthermore,we utilized Western blotting to analyze senescenceassociated proteins p21 and proliferating cell nuclear antigen(PCNA),thereby evaluating the role of FEN1 in cellular senescence.The impact of FEN1 on cisplatin sensitivity was investigated via the CCK-8 cell counting assay.Additionally,we investigated how FEN1 inhibitors might impact NB cell proliferation and enhance the therapeutic efficacy of cisplatin treatment.Results:FEN1 was found to be highly expressed in stage IV NB and showed a strong association with cisplatin sensitivity,establishing it as a critical molecular marker linked to poor patient prognosis.Notably,elevated FEN1 expression correlated with reduced sensitivity to cisplatin,as evidenced by higher IC50 values.In the SH-SY5Y cell line,FEN1 knockdown led to significant reductions in cell proliferation,invasion,and migration,along with an increase inβ-galactosidase staining—indicative of senescence.This knockdown also resulted in elevated levels of the p21 protein and decreased expression of PCNA,concurrently lowering cisplatin IC50 values.Conversely,FEN1 overexpression in the SK-N-SH cell line resulted in enhanced cell proliferation,invasion,and migration.This overexpression was associated with reducedβ-galactosidase staining,decreased levels of p21,and increased expression of PCNA,ultimately resulting in higher cisplatin IC50 values.Importantly,FEN1 inhibitors alone significantly impeded NB cell proliferation,and their combination with cisplatin further amplified this inhibitory effect compared to cisplatin treatment alone.Conclusions:Bioinformatics and sequencing analyses indicate that the senescence-related gene FEN1 is significantly associated with cisplatin sensitivity and adverse prognosis in pediatric NB.FEN1 plays a pivotal role in regulating NB cell proliferation,invasion,and migration,thereby facilitating cancer progression.Furthermore,it influences cisplatin sensitivity through its effects on cellular senescence.FEN1 inhibitors demonstrate potential both as monotherapies and in conjunction with cisplatin,suggesting that targeting FEN1 may be represent a valuable strategy for improving outcomes in high-risk NB patients.
文摘BACKGROUND Neuroblastoma(NB)is one of the most common malignancies in children.Metastasis in NB is not uncommon.However,nasal metastases are rare.Here,we reported two pediatric cases of nasal metastases.CASE SUMMARY Case 1 was a 3-year-old boy without a history of NB.Case 2 was a 10-year-old girl who had a history of NB for 6 years.Both of them presented with symptoms of nasal and sinus masses such as epistaxis or discharge from the nose.The radiologic imaging results revealed masses in the nasal cavity or nasopharynx in both cases and a mass in the right adrenal gland of case 1.The pathologic examination of biopsy samples of their nasal masses revealed“small round bluecell tumor”along with abundant vascular fibrous septa.The tumor cells expressed synaptophysin,cluster of differentiation 56,chromogranin A,paired like homeobox protein 2B and a very high Ki67 index in both case but were negative for vimentin,desmin,leucocyte common antigen and cytokeratin.Myelocytomatosis viral related oncogene,neuroblastoma derived(MYCN)amplification was detected in both cases.Finally,the two cases were diagnosed as nasal metastases from NB based on the clinical and pathologic findings.The two patients affected by NB were>18 mo old,the primary tumor location was adrenal gland,and they presented with multiple metastases.CONCLUSION It is difficult to differentiate between metastatic NB in the nose and olfactory neuroblastoma in the absence of a history of NB.Paired like homeobox protein 2B can play an important role in the diagnosis and differential diagnosis of this disease.
文摘Esthesioneuroblastoma is a malignant tumor, arising in the upper nasal cavity, that could spread to the frontal lobe of the brain as well as metastasize to the lymph nodes. Due to the low incidence of this tumor, FDA-approved treatment modalities do not exist and clinical trials have not been performed. We present an interesting case of a 66-year-old female, diagnosed with Kadish stage B esthesioneuroblastoma and stage IIA nonsmall cell carcinoma of the lung, who benefited from our treatment. Both malignancies were diagnosed in 2002 at which time the patient consented to undergo left upper lobectomy for her lung cancer, but she refused the craniofacial resection and radiation therapy recommended for treatment of her esthesioneuroblastoma. From 2003 to 2004 she received treatment at the Burzynski Clinic with oral sodium phenylbutyrate (0.2 g/kg/day). She tolerated the treatment very well without significant adverse events. Gradual reduction in her tumor size was confirmed by repeat MRIs. From treatment start in March 2003 to December 2003 her tumor decreased by 40%. Subsequent MRI from March 2004 revealed increased tumor size, which, however, was still a 13% reduction from the baseline MRI. What is important to mention is that in addition to shrinkage of the esthesioneuroblastoma, the patient obtained the clinical benefit of 3.5-years longer survival than was predicted for her lung cancer—whereas the median survival for a patient with stage IIA adenocarcinoma of the left upper lobe of the lung is approximately two years, our patient survived more than five and a half years. The effect of phenylbutyrate (PB) and its metabolite phenylacetate on neuroblastoma and lung cancer is documented by numerous preclinical studies and is also evident in this case. It is proposed that the activity of these two compounds is mediated through increased expression of the p21 tumor suppressor gene. p21 is a strong inhibitor of cyclin-D and cyclin-dependent kinase 4, which contribute to undifferentiated phenotype in neuroblastoma and are instrumental in cell cycle progression from G1 to S phase. It is hoped that future research and combination of PB with other chemotherapeutic and targeted agents will provide better control of esthesioneuroblastoma and lung cancer.
文摘Olfactory neuroblastoma (esthesioneuroblastoma, ЕNB) is a rare tumor arising from the olfactory neuroepithelium. We report a case of ЕNB located in inferior nasal concha, combined with thyroid gland carcinoma and gastrointestinal stromal carcinoma in a 77-year-old man. The tumor was resected endonasally. When the final diagnosis of olfactory neuroblastoma was confirmed by histopathologic examination and immunohistochemical staining, the PET/CT examination was performed. The imaging revealed a small focus of a moderately increased cancer activity in the thyroid region. A gastrointestinal stromal carcinoma was detected one year after the resection of the thyroid gland. We discuss the clinical appearance of ENB, staging systems, diagnosis and management. During the endonasal surgery, ENB was removed entirely. Seven days after operation, in order to monitor the postoperative result, PET/CT was performed and a papillary thyroid cancer was detected. One year after the thyroid surgery, gastroendoscopy showed a neoplastic formation in the stomach. In conclusion, we state that when identified as aggressive tumors such as ENB, it is necessary to provide regular examinations in order to detect distant ENB metastases or other neoplastic localisations.
文摘This article presents a case of a patient with relapsed esthesioneuroblastoma (ENB), an aggressive rare tumor that arises from the specialized sensory epithelial olfactory cells in the skull base area, which was initially treated with endoscopic surgery, followed by adjuvant radiotherapy. After local relapse, new surgical approaches and subsequent lines of platin-based chemotherapy were performed. A PET-CT with <sup>68</sup>GALIUM DOTATATOC (PET-DOTATOC) showed intense uptake of disease, compatible with the presence of somatostatin receptors, in the face, nodes, liver, bones, and meningeal area. Treatment with 4 cycles of <sup>177</sup>Lutetium-Dotatate was performed, followed by maintenance octreotide, with a major radiological and clinical response that is lasting more than 1 year after treatment. This article describes a rare case of a skull-base tumor, with multiple recurrences, in which disease control was achieved with a targeted Peptide Receptor Radionuclide Therapy (PRRT) with <sup>177</sup>Lutetium-Dotatate, and discusses factors that could influence the incorporation of this form of therapy. Previous case reports proved the potential efficacy of this therapy usually given for low-grade neuroendocrine tumors and will be carefully reviewed.
基金Supported by the National Natural Science Foundation of China with Grant No.ul204815
文摘Objective:To evaluate the efficacy of allicin combined with cyclophosphamide on neuroblastoma(NB)-bearing mice and explore the immunological mechanism in it.Methods:A total of 30NB-bearing mice were equally randomized into model group,cyclophosphamide group and combined therapy group,10 nudemice were set as normal saline(NS) group.Cyclophosphamide group and combined therapy group were weekly injected with 60 mg/kg cyclophosphamide for four weeks;besides,combined therapy group was given with allicin(10 mg/kg/d) by gastric perfusion for 4 weeks;model group and NS group were given with the same volume of NS.Serum VEGF content was detected by ELISA pre-treating(0 d) and on the 3rd d.14 th d and 28 th d;on29th d,all mice were sacrificed and the tumor,liver,spleen and thymic tissues were weighted.Tumors were made into paraffin section for detecting tumor cell apoptosis and proliferation by TUNEI.and BrdU method,respectively.Survival curves were drawn by Kaplan-Meier method.Results:After treatment,both treatment groups relieved on viscera indexes,VEGF level,T cell subsets distribution and tumor growth and each index of combined therapy group was better than cyclophosphamide group(P【0.05 or 0.01);only combined therapy group could significantly increase the lifetime of NB-bearing mice(χ~2=5.667,P=0.017).Conclusions:Allicin can improve T cell subsets distribution and inhibit VEGF expression through its immunomodulatory activity,thereby improve the efficiency on NB in coordination with cyclophosphamide.
文摘Neuroblastoma(NB) is the most common extracranial solid tumor in children. Diarrheal NB is quite rare and is not easy to diagnose in the early stage. Six cases of diarrheal NB in our hospital treated from 1996 to 2006 were retrospectively analyzed, including characteristics such as electrolyte imbalance, pathologic features, vasoactive intestinal peptide(VIP) immunohistochemical staining results, treatment, and prognosis. All patients were boys with 3-8 loose or watery stools each day and routine fecal tests were normal. Abdominal tumors were identified by B-ultrasound. Drugs were ineffective. Three patients underwent surgery, and the remaining three patients received surgery and chemotherapy. Diarrhea stopped after treatment in five patients. Two patients died due to intractable hypokalemia. The tumor was located in the adrenal gland in four patients, in the upper retroperitoneum in one patient, and in the presacral area in one patient. Pathologic findings were NB and ganglioneuroblastoma. Five patients were at clinical stage Ⅰ-Ⅱ, and one was at stage Ⅲ. Four patients survived(followed-up for 6 mo to 4 years). Immunohistochemical staining for VIP was positive. Refractory diarrhea is a paraneoplastic syndrome of NB and is rare. Patients aged 1-3 years who present with chronic intractable diarrhea should be followed closely. Intractable diarrhea, hypokalemia, and dysplasia are the initial clinical manifestations. Increased VIP is characteristic of this disease. Potassium supplementation plays a vital role in the treatment procedure, especially preoperatively. The prognosis of diarrheal NB is good following appropriate treatment.
文摘Introduction: Brain metastasis is common in relapsed neuroblastoma patients, but the characteristics of brain metastasis remain largely unknown. This study aimed to investigate the status of brain metastasis with neuroblastoma in South China.Methods: In this retrospective case?based study, 106 patients with stage 4 neuroblastoma from the Department of Pediatric Oncology in Sun Yat?sen University Cancer Center between January 2004 and May 2013 were included. The incidence, risk factors, and survival status of these patients were reviewed and analyzed.Results: Of the 106 patients, 11(10.4%) developed brain metastasis, accounting for 20.0% of 55 patients with relapse or progression. The age at initial diagnosis of the 11 patients ranged from 2 to 10 years(median 4 years), which was younger than that of the patients without brain metastasis(median 5 years, range 1–10 years, P = 0.073). The male to female ratio of the 11 patients was 8:3, which was not signiicantly diferent from that of the patients with?out brain metastasis(P = 0.86). Patients with brain metastasis had higher lactate dehydrogenase levels than those without brain metastasis, but the diferences were not signiicant(P initial diagnosis to the develo= 0.076). Eight patients died, and 3 patients survived. The median interval from thepment of brain metastasis was 18 months(range 6–32 months). The median survival was 4 months(range 1 day to 29 months) after the diagnosis of brain metastasis. The median interval from the manifestation of brain metastasis to death was 3 months(range 1 day to 11 months).Conclusions: High?risk factors for brain metastasis in cases of neuroblastoma include bone marrow involvement and a younger age at initial diagnosis. Nevertheless, multiple treatment modalities can improve disease?free survival.
文摘Thymoquinone(TQ), an active component derived from the medial plant Nigella sativa, has been used for medical purposes for more than 2 000 years. Recent studies have reported that TQ blocked angiogenesis in animal model and reduced migration, adhesion, and invasion of glioblastoma cells. We have recently shown that TQ could exhibit a potent cytotoxic effect and induce apoptosis in mouse neuroblastoma(Neuro-2a) cells. In the present study, TQ treatment markedly decreased the adhesion and migration of Neuro-2a cells. TQ down-regulated MMP-2 and MMP-9 protein expression and m RNA levels and their activities. Furthermore, TQ significantly down-regulated the protein expression of transcription factor NF-κB(p65) but not significantly altered the expression of N-Myc. Taken together, our data indicated that TQ's inhibitory effect on the migration of Neuro-2a cells was mediated through the suppression of MMP-2 and MMP-9 expression, suggesting that TQ treatment can be a promising therapeutic strategy for human malignant neuroblastoma.
基金supported by the National Natural Science Foundation of China,No.81272986the Natural Science Foundation of Shandong Province,No.ZR2011HZ002
文摘This study investigated the effects of small interfering RNA （siRNA）-mediated silencing of chemokine receptor 4 （CXCR4） on the invasion capacity of human neuroblastoma cell line SH-SY5Y in vitro. Three siRNAs targeting CXCR4 were chemically synthesized and individually transfected into SH-SY5Y cells. Expression of CXCR4 mRNA and protein was signiifcantly sup-pressed in transfected cells by all three sequence-speciifc siRNAs compared with control groups. Furthermore, the invasion capacity of SH-SY5Y cells was signiifcantly decreased following trans-fection with CXCR4-speciifc siRNA compared with the control groups. These data demonstrate that down-regulation of CXCR4 can inhibit in vitro invasion of neuroblastoma.
基金the National Natural Sciences Foundation of China(39470739)the Ministry of Public Health Research Foundation(20122167)the Doctor Startup-Natural Science Foundation of Li-aoning Province (20041047)
文摘Objective To study the effect of γ-interferon （IFNγ）, tumor necrosis factor related apoptosis inducing ligand （TRAIL）, and cisplatin or etoposide induced apoptosis in human neuroblastoma cell line SH-SY5Y and its possible molecular mechanisms. Methods The expressions of Caspase 8 mRNA and protein were detected with RT-PCR and Western blot analysis. The effects of IFNγ, TRAIL, IFNγ ＋ TRAIL, IFNγ ＋ Caspase 8 inhibitor ＋ TRAIL, IFNγ ＋ cisplatin ＋ TRAIL, and IFNγ ＋ etoposide ＋ TRAIL on the growth and apoptosis of SH-SY5Y cells were detected with the methods of MTT and flow cytometry. The relative Caspase 8 activity was measured with colorimetric assay. Results Caspase 8 was undetectable in SH-SY5Y cells but an increased expression of Caspase 8 mRNA and protein was found after treatment with IFNγ. SH-SY5Y ceils themselves were not sensitive to TRAIL, but those expressing Caspase 8 after treatment with IFNγ were. The killing effect of TRAIL on SH-SY5Y cells expressing Caspase 8 was depressed by Caspase 8 inhibitor. Cisplatin and etoposide could enhance the sensitivity of TRAIL on SH-SY5Y cells. The relative Caspase 8 activity of SH-SY5Y cells in IFNγ ＋ TRAIL group was significantly higher than those of control group, IFNγ group, TRAIL group, and inhibitor group （ P 〈 0. 01 ）. There was no significant difference among IFNγ ＋ TRAIL group, IFNγ ＋ cisplatin ＋ TRAIL group, and IFNγ ＋ etoposide ＋ TRAIL group. Conclusions IFNγ could sensitize SH-SY5Y cells to TRAIL-induced apoptosis and this may be realized by the up-regulation of Caspase 8. Cisplatin and etoposide could enhance the killing effect of TRAIL on SH-SY5Y cells.
基金supported by grants from the National Natural Science Foundation of China(No.82002635,82002636and 82173593)GuangzhouScienceand TechnologyProject(No.202102021227 and202102020421)+1 种基金the Science Technology and Innovation Commission of Shenzhen(No.JCYJ20220531093213030)Guangzhou Municipal Basic Research Program Joint Funding of City and Hospitals(No.202201020622).
文摘Objective:AlkB homolog 5(ALKBH5)has been proven to be closely related to tumors.However,the role and molecular mechanism of ALKBH5 in neuroblastomas have rarely been reported.Methods:The potential functional single-nucleotide polymorphisms(SNPs)in ALKBH5 were identified by National Center for Biotechnology Information(NCBI)dbSNP screening and SNPinfo software.TaqMan probes were used for genotyping.A multiple logistic regression model was used to evaluate the effects of different SNP loci on the risk of neuroblastoma.The expression of ALKBH5 in neuroblastoma was evaluated by Western blotting and immunohistochemistry(IHC).Cell counting kit-8(CCK-8),plate colony formation and 5-ethynyl-2'-deoxyuridine(EdU)incorporation assays were used to evaluate cell proliferation.Wound healing and Transwell assays were used to compare cell migration and invasion.Thermodynamic modelling was performed to predict the ability of miRNAs to bind to ALKBH5 with the rs8400 G/A polymorphism.RNA sequencing,N6-methyladenosine(mA)sequencing,mA methylated RNA immunoprecipitation(MeRIP)and a luciferase assay were used to identify the targeting effect of ALKBH5 on SPP1.Results:ALKBH5 was highly expressed in neuroblastoma.Knocking down ALKBH5 inhibited the proliferation,migration and invasion of cancer cells.miR-186-3p negatively regulates the expression of ALKBH5,and this ability is affected by the rs8400 polymorphism.When the G nucleotide was mutated to A,the ability of miR-186-3p to bind to the 3'-UTR of ALKBH5 decreased,resulting in upregulation of ALKBH5.SPPI is the downstream target gene of the ALKBH5 oncogene.Knocking down SPP1 partially restored the inhibitory effect of ALKBH5 downregulation on neuroblastoma.Downregulation of ALKBH5 can improve the therapeutic efficacy of carboplatin and etoposide in neuroblastoma.Conclusions:We first found that the rs8400 G>A polymorphism in the m6A demethylase-encoding gene ALKBH5 increases neuroblastoma susceptibility and determines the related mechanisms.The aberrant regulation of ALKBH5 by miR-186-3p caused by this genetic variation in ALKBH5 promotes the occurrence and development of neuroblastoma through the ALKBH5-SPP1 axis.
基金supported by the National Natural Science Foundation of China (No.30870507)partially supported by a grant from the Ministry of Science and Technology of China (No.2005CB522603)
文摘Objective： Id2 is a natural inhibitor of the basic helix-loop-helix（bHLH） transcription factors. Although it is well known that active ld2 prevents differentiation and promotes cell cycle progression and tumorigenesis, the molecular events that regulate Id2 activity remain to be investigated. Methods： Yeast two-hybrid, mammalian two-hybrid, GST-pulldown and immunoprecipitation （ColP） assays were used to screen and identify novel Id2 interactors. Luciferase assays were used to detect E47-mediated transcription activity. Colony formation and BrdU incorporation assays were used to determine cellular proliferation abilities. Northorn blot, western blot and quantitative PCR methods were used to measure gene expression levels. Electrophoretic mobility shift assays （EMSAs） were performed to investigate protein/DNA binding. Results： The LIM-only protein FHL2 （four-and-a-half-LIM-only protein 2） was identified to be a novel Id2 interactor. The HLH domain within Id2 is not required for its interaction with FHL2. FHL2 antagonizes the inhibitory effect of Id2 on the basic helix-loop-helix protein E47-mediated transcription. FHL2 prevents the formation of Id2-E47 heterdimer, thus releasing E47 to its target DNA and restoring its transcriptional activity. FHL2 expression was remarkably up-regulated during retinoic acid-induced differentiation of neuroblastoma cells, during which the expression of Id2 is opposite to that. Ectopic FHL2 expression in neuroblastoma cells markedly reduces the transcriptional and cell-cycle promoting functions of Id2. Conclusion： These results indicate that FHL2 is an important repressor of the oncogenic activity of Id2 in neuroblastoma cells.
基金supported by the Research and Development Project Fund of Science and Technology Plan Program of Science and Technology Bureau of Beijing of China,No.Z111107067311022
文摘The active ingredient of ginseng,ginsenosides Rg1,has been shown to scavenge free radicals and improve antioxidant capacity.This study hypothesized that ginsenosides Rg1 has a protective role in human neuroblastoma cells injured by H2O2.Ginsenosides Rg1 at different concentrations（50 and 100 μM） was used to treat H2O2（150 μM）-injured SH-SY5 Y cells.Results demonstrated that ginsenoside Rg1 elevated the survival rate of SH-SY5 Y cells injured by H2O2,diminished the amount of leaked lactate dehydrogenase,and increased superoxide dismutase activity.Ginsenoside Rg1 effectively suppressed caspase-3 immunoreactivity,and contributed to heat shock protein 70 gene expression,in a dose-dependent manner.These results indicate that ginsenoside Rg1 has protective effects on SH-SY5 Y cells injured by H2O2 and that its mechanism of action is associated with anti-oxidation and the inhibition of apoptosis.
文摘Most therapeutic protocols for child cancers use cytotoxic agents which have a narrow therapeutic index,and resulting in severe acute and chronic toxicities to normal tissues. Despite the fact that most child cancer patients achieve complete remission after chemotherapy,death still occurs due to relapse of persistent minimal residual disease(MRD) which remaining after initial cytotoxic chemotherapy. Advanced neuroblastoma(NB) is a leading cause of cancer deaths in young children. Retinoids are an important component of advanced NB therapy at the stage of MRD,yet half of all patients treated with 13-cis-retinoic acid still relapse and die. More effective combination therapies,with a lower side-effect profile,are required to improve outcomes for NB. Fenretinide or N-4-hydroxyphenyl retinamide is a synthetic derivative of retinoic acid which works on cancer cells through nuclear receptor-dependent and-independent signalling mechanisms. Moreover,several histone deacetylase inhibitors have entered early phase trials,and,suberoylanilide hydroxamic acid has been approved for use in adult cutaneous T cell lymphoma. A number of studies suggest that retinoid signal activation is necessary for histone deacetylase inhibitor activity. A better understanding of their mechanism of actions will lead to more evidence-based retinoid combination therapies.
文摘Cancers are characterized by deregulation of multiple signaling pathways and thus monotherapies are hardly effective. Neuroblastoma, which often occurs in adrenal glands, is the most common childhood malignancy. Malignant neuroblastoma resists traditional treatments and further studies are needed for effective therapeutic interventions. We evaluated synergistic efficacy of N-(4-hydroxyphenyl) retinamide (4-HPR) and genistein (GST) for induction of apoptosis in human malignant neuroblastoma SH-SY5Y and SK-N-BE2 cells in culture and activation of multiple pathways for increasing apoptosis in ectopic and orthotopic neuroblastoma xenografts in nude mice. Combination of 4-HPR and GST synergistically reduced cell viability, caused subG1 accumulation, increased caspase-3 activity for apoptosis in vitro and reduced tumor growth in vivo. Western blotting indicated that combination therapy down regulated Id2 to induce differentiation, increased pro-apoptotic Bax and decreased anti-apoptotic Bcl-2 leading to an increase in Bax:Bcl-2 ratio, increased mitochondrial Bax level, caused mitochondrial release of Smac/Diablo, down regulation of the baculovirus inhibitor-of-apoptosis repeat containing (BIRC) proteins such as BIRC-2 and BIRC-3, and activation of calpain and caspase-3 in SH-SY5Y xenografts. Accumulation of apoptosis-inducing-factor (AIF) in cytosol and increase in caspase-4 activation suggested involvement of mitochondrial pathway and endoplasmic reticulum (ER) stress, respectively, for apoptosis in SH-SY5Y xenografts. In situ immunofluorescent labelings of SH-SY5Y and SK-N-BE2 xenograft sections showed overexpression of calpain, caspase-12, and caspase-3, and AIF, suggesting induction of mitochondrial caspase-dependent and caspase-independent pathways for apoptosis. Collectively, synergistic effects of 4-HPR and GST induced mitochondrial pathways and also ER stress for increasing apoptosis in ectopic and orthotopic neuroblastoma xenografts in nude mice.
文摘The effects of tissue factor （TF） on doxorubicin-induced apoptosis in human neuroblastoma were investigated. The expression of TF was examined by Western blotting. TFsiRNA-pSUPER plasmid was constructed by inserting specific 19-nt silencing sequence targeting TF gene into pSUPER vector. Transfection of TFsiRNA-pSUPER was performed using lipofectamine^2000. The cytotoxicity of doxorubicin was determined by WST assay. The activation of Caspase-3 and PARP induced by doxorubicin was tested by Western blotting. The apoptotic cells were stained by Hochest33342 and counted under fluorescence inverted microscope. It was found that human neuroblastoma cell line SK-N-MC expressed high level of TE Knockdown of the TF expression was achieved by transfection of TFsiRNA-pSUPER on SK-N-MC cells in a dose-dependent manner. Inhibition of TF significantly decreased the viability of transfected SK-N-MC cells treated with different concentrations of doxorubicin. Cleavage of Caspase-3 and PARP was enhanced in transfected SK-N-MC cells with down-regulation of TF. TFsiRNA treatment significantly increased the number of apoptotic cells in transfected SK-N-MC cells as compared with those control cells （P〈0.05） when these cells were exposed to 1 μg/mL doxorubicin for 8 h. These results suggested that knockdown of the TF expression by specific siRNA vector could increase the cytotoxicity of doxorubicin and enhance doxorubicin-induced apoptosis in human neuroblastoma cells. Over-expression of TF might contribute to chemotherapy resistance in human neuroblastoma and its progression, at lest in part, by regulating doxorubicin-induced apoptosis.
