Stage IV-S Neuroblastoma Research Articles

Identification of different ALK mutations in a pair of neuroblastoma cell lines established at diagnosis and relapse.

Anaplastic Lymphoma Kinase (ALK) is a transmembrane receptor kinase that belongs to the insulin receptor superfamily and has previously been shown to play a role in cell proliferation, migration and invasion in neuroblastoma. Activating ALK mutations are reported in both hereditary and sporadic neuroblastoma tumours, and several ALK inhibitors are currently under clinical evaluation as novel treatments for neuroblastoma. Overall, mutations at codons F1174, R1275 and F1245 together account for ∼85% of reported ALK mutations in neuroblastoma. NBLW and NBLW-R are paired cell lines originally derived from an infant with metastatic MYCN amplified Stage IVS (Evans Criteria) neuroblastoma, at diagnosis and relapse, respectively. Using both Sanger and targeted deep sequencing, this study describes the identification of distinct ALK mutations in these paired cell lines, including the rare R1275L mutation, which has not previously been reported in a neuroblastoma cell line. Analysis of the sensitivity of NBLW and NBLW-R cells to a panel of ALK inhibitors (TAE-684, Crizotinib, Alectinib and Lorlatinib) revealed differences between the paired cell lines, and overall NBLW-R cells with the F1174L mutation were more resistant to ALK inhibitor induced apoptosis compared with NBLW cells. This pair of cell lines represents a valuable pre-clinical model of clonal evolution of ALK mutations associated with neuroblastoma progression.

A Novel Anticancer Agent, 8-Methoxypyrimido[4',5':4,5]thieno(2,3-b) Quinoline-4(3H)-One Induces Neuro 2a Neuroblastoma Cell Death through p53-Dependent, Caspase-Dependent and -Independent Apoptotic Pathways.

Neuroblastoma is the most common cancer in infants and fourth most common cancer in children. Despite recent advances in cancer treatments, the prognosis of stage-IV neuroblastoma patients continues to be dismal which warrant new pharmacotherapy. A novel tetracyclic condensed quinoline compound, 8-methoxypyrimido [4′,5′:4,5]thieno(2,3-b) quinoline-4(3H)-one (MPTQ) is a structural analogue of an anticancer drug ellipticine and has been reported to posses anticancer property. Study on MPTQ on neuroblastoma cells is very limited and mechanisms related to its cytotoxicity on neuroblastoma cells are completely unknown. Here, we evaluated the anticancer property of MPTQ on mouse neuro 2a and human SH-SY5Y neuroblastoma cells and investigated the mechanisms underlying MPTQ-mediated neuro 2a cell death. MPTQ-mediated neuro 2a and SH-SY5Y cell deaths were found to be dose and time dependent. Moreover, MPTQ induced cell death reached approximately 99.8% and 90% in neuro 2a and SH-SY5Y cells respectively. Nuclear oligonucleosomal DNA fragmentation and Terminal dUTP Nick End Labelling assays indicated MPTQ-mediated neuro 2a cell death involved apoptosis. MPTQ-mediated apoptosis is associated with increased phosphorylation of p53 at Ser15 and Ser20 which correlates with the hyperphosphorylation of Ataxia-Telangiectasia mutated protein (ATM). Immunocytochemical analysis demonstrated the increased level of Bax protein in MPTQ treated neuro 2a cells. MPTQ-mediated apoptosis is also associated with increased activation of caspase-9, -3 and -7 but not caspase-2 and -8. Furthermore, increased level of caspase-3 and cleaved Poly (ADP Ribose) polymerase were observed in the nucleus of MPTQ treated neuro 2a cells, suggesting the involvement of caspase-dependent intrinsic but not extrinsic apoptotic pathway. Increased nuclear translocation of apoptosis inducing factor suggests additional involvement of caspase-independent apoptosis pathway in MPTQ treated neuro 2a cells. Collectively, MPTQ-induced neuro 2a cell death is mediated by ATM and p53 activation, and Bax-mediated activation of caspase-dependent and caspase-independent mitochondrial apoptosis pathways.

Abdominal Compartment Syndrome Due to Liver Metastases of Stage IV-S Neuroblastoma - is Liver Transplantation a Reasonable Option?

Background: Stage IV-S neuroblastoma in infancy has a good prognosis with a high rate of complete remission. Some patients develop extended hepatic metastases with the risk of an abdominal compartment syndrome and subsequent liver failure. Liver transplantation as a life saving procedere has not been established yet. Case report: An abdominal mass led to the diagnosis of a neuroblastoma of the right glandula adrenalis in a six week-old boy. With the evidence of liver metastases and bone marrow infiltration less than 1% the diagnosis of a stage IV-S neuroblastoma was made. Because of the expected good prognosis a chemotherapy was not started immediately. In the following time the child devoloped extensive liver metastases with consequent abdominal compartment syndrome. A chemotherapy was started and at the same time several abdominal extensions with abdominal patches were performed. Nevertheless, the tumor size did not decrease and the patient developed an acute liver failure due to impaired liver perfusion. Because of acute deterioration of liver function with coagulopathy and hyperammonemia the patient was listed for emergency liver transplantation. A further course of chemotherapy was declined because the histology of the liver did only show few vital tumor cells but in contrast extended liver necrosis. The child received a liver transplant (left lateral split of a deceased donor, graft weight 550 g; graft-to-recipient weight-ratio: 11%) at the age of 3 months (body weight: 5 kg); the adrenal neuroblastoma was removed simultaneously. Because of the large-for-size situation there was a continuous impaired transplant perfusion leading to re-transplantation (left lateral split of a child, graft weight: 250 g; graft-to-recipient weight-ratio: 5%). The postoperative course was complicated by a very delayed abdominal closure and recurrent infections. The patient has a good liver function 3 months after liver transplantation without tumor recurrence and without necessity of further chemotherapy. Conclusion: Liver transplantation can be a life-saving option for infants with stage IV-S neuroblastoma and liver failure due to abdominal compartment syndrome. Because of the general good prognosis of this tumor, the need of a post-operative chemotherapy has to be discussed when the primary neuroblastoma is removed simultaneously during transplantation procedere. Problems can occur as a result of skin retraction during prolonged abdominal closure.

Anaphylactic Reaction Owing to Ondansetron Administration in a Child With Neuroblastoma and Safe Use of Granisetron: A Case Report

A 1-year-old girl with stage-IV neuroblastoma developed ondansetron hydrochloride anaphylaxis. Safe use of granisetron as an antiemetic after skin prick, oral, and intravenous challenge tests is presented. We present this case to emphasize that ondansetron hydrochloride may cause a serious anaphylactic reaction. In such a case, granisetron may be given to patients as an antiemetic after some provocative tests performed.

99mTc-HYNIC octreotide in neuroblastoma

Disease status assessment of neuroblastoma patients requires computed tomography (or magnetic resonance imaging), bone scan, metaiodobenzylguanidine (MIBG) scan, bone marrow tests, and urine catecholamine measurements. There is no clinical experience concerning the evaluation of these patients by means of technetium-99m (99mTc)-somatostatin analog scintigraphy. Furthermore, these radiopharmaceuticals are promising imaging agents owing to their lower cost, availability, dosimetry, and ease of preparation. An 8-year-old boy already diagnosed with stage-IV neuroblastoma received chemotherapy. In the follow-up, after obtaining the parents' informed consent, iodin 131 (131I)-MIBG and 99mTc-6-hydrazinopyridine-3-carboxylic acid (HYNIC)-octreotide scans were done on separate days to evaluate tumor extension. Even as the 131I-IBG scan showed mild diffuse uptake in the projection of both lung hili, the 99mTc-HYNIC-octreotide scan showed multiple axial and appendicular bone uptakes and paravertebral, abdominal, mediastinal, and supraclavicular ganglionar uptakes. The 99mTc-HYNIC-octreotide showed much more lesion extension than the 131I-MIBG. Therefore, 99mTc-HYNIC-octreotide may be a promising radiopharmaceutical for the evaluation of neuroblastoma patients. This finding justifies the preliminary evaluation of this tracer in the context of a clinical trial.

Identification of DEIN, a Novel Gene with High Expression Levels in Stage IVS Neuroblastoma

Neuroblastoma at stage IVS, defined by dissemination to specific tissues and age <1 year at diagnosis, regularly follows spontaneous regression without cytotoxic treatment. To uncover the molecular characteristics of this subtype, Serial Analysis of Gene Expression (SAGE) profiles from stage IVS and fatal stage IV tumors were compared. A SAGE tag (GCAACTTAAC) was detected that was overrepresented in stage IVS disease and that had no reliable match in current National Center for Biotechnology Information databases of SAGE profiles, thus pointing to a novel gene. The corresponding gene, which maps to chromosome 4q33-34, was identified using a modified 3'- and 5'-rapid amplification of cDNA ends PCR and was designated as DEIN (differentially expressed in neuroblastoma). The gene comprises five transcript variants and its sequence overlaps with expressed sequences of the yet uncharacterized UniGene cluster Hs.61435. DEIN exhibits nucleotide sequence conservation over a broad range of species with an overall homology of 65% between human and mouse. As none of the predicted amino acid sequences is homologous to known proteins, it remains to be determined whether DEIN represents a coding or noncoding RNA. Northern blot analysis and semiquantitative reverse transcription-PCR showed high DEIN expression in neuroblastoma, whereas expression was absent or weak in most normal adult tissues. Analysis of 121 primary neuroblastomas by real-time reverse transcription-PCR revealed a strong association with age at diagnosis <1 year and particularly with stage IVS disease (both P < 0.001). The characteristic expression pattern of DEIN suggests a specific role of this gene in the unique biology of stage IVS tumors and may help to molecularly define this special subtype of neuroblastoma.

Continuous veno-venous haemodiafiltration in an infant with a stage IVs neuroblastoma and acute renal failure

Continuous veno-venous haemodiafiltration in an infant with a stage IVs neuroblastoma and acute renal failure

Rasburicase for the Management of Tumor Lysis Syndrome in Neonates

To describe the management of tumor lysis syndrome (TLS) with rasburicase in 2 patients who presented with cancer within the first month of life and compare and contrast both cases with respect to their underlying renal physiology, management, and eventual outcome. TLS developed in 2 neonates born at 38 weeks' gestational age; both were managed in part with rasburicase. One patient was a 21-day-old infant who received 2 days of induction chemotherapy for the treatment of congenital Stage IV-S neuroblastoma. With a single 0.2 mg/kg dose of rasburicase, the serum urate level normalized and the infant completed therapy without incident. The second patient was a 4-day-old neonate with congenital precursor-B cell acute lymphoblastic leukemia who presented with spontaneous TLS complicated by renal dysfunction. Despite several doses of intravenous rasburicase (2 doses of 0.1 mg/kg and 4 doses of 0.2 mg/kg), as well as aggressive supportive therapy, the infant died of complications arising from uncontrolled TLS. Neonates may be at particular risk for TLS given their immature renal function and its predisposition toward metabolic derangements. While rasburicase has the potential to provide a rapid reversal of TLS in this patient population, when TLS is complicated by pre-existing acute renal failure, additional interventions and alternative anti-tumor strategies may be necessary for a successful outcome. When managing TLS in infancy, clinicians must consider the relative degree of renal immaturity and its predisposition toward metabolic derangements. Rasburicase appears to be well tolerated and effective in lowering serum urate concentrations in the treatment of therapy-related TLS in neonates. However, in instances of spontaneous TLS complicated by the normally low glomerular filtration rate in the newborn infant, the use of rasburicase and other supportive care measures may still be inadequate, warranting further study.

Transient Increase of Serum Granulysin in a Stage IVs Neuroblastoma Patient during Spontaneous Regression: Case Report

Stage IVs neuroblastoma is a unique cancer that shows spontaneous regression, although it does exhibit metastasis to the spleen or liver [1]. The mechanisms of spontaneous regression of the tumor are not precisely known, but endogenously programmed biological mechanisms such as apoptosis are suspected [2]. In addition, immunologic mechanisms such as cell-mediated immunity have also been speculated [3]. Granulysin is a recently defined cytolytic granule protein of natural killer cells and cytotoxic T-lymphocytes [4,5]. It has been demonstrated to possess antimicrobial effects when used in isolation against a variety of pathogens, including bacteria, fungi, and parasites [6-8]. Furthermore, granulysin is believed to contribute to antitumor and antiviral immune responses through its tumoricidal and antiviral activities [9]. Recently, we have established an enzymelinked immunosorbent assay system that detects granulysin in the serum [5]. We treated a girl with stage IVs neuroblastoma; during the spontaneous regression of the tumor, her serum granulysin level was markedly increased. A 2-month-old girl was admitted to our hospital because of an abdominal tumor. She had been admitted to a hospital at 1 month of age because of bronchiolitis from a respiratory syncytial virus infection; hepatomegaly was not noted at that time. The parents noticed distention in the patient’s abdomen, which was progressing day by day 1 week before admission. A computed tomography scan showed a markedly enlarged liver, which extended to the pelvic cavity. A systemic bone survey revealed no abnormal findings. Magnetic resonance imaging showed heterogeneous multifocal masses occupying the liver, but the primary site could not be determined. Scintigraphy testing (metaiodobenzylguanidine I 123) disclosed no metastatic site other than the liver. Bone marrow aspirated from the right tibial bone showed no invaTransient Increase of Serum Granulysin in a Stage IVs Neuroblastoma Patient during Spontaneous Regression: Case Report

Pediatric Renal Carcinoma Associated with Xp11.2 Translocations/TFE3 Gene Fusions and Clinicopathologic Associations

Renal cell carcinomas (RCCs) are rare in children and studies of their subtypes and clinicopathologic associations are limited to small series. We identified 8 patients with RCC treated at our institution between 1981 and 2003, reviewed their clinicopathologic features, cytogenetics findings, and evaluated the status of TFE3 expression by immunohistochemistry and numerical chromosomal alterations by interphase fluorescent in situ hybridization on paraffin-embedded tissue. These 8 patients (5 female and 3 male) had diploidy, and 5 had morphologic features compatible with the recently described RCC associated with Xp11.2 translocations/TFE3 gene fusions and demonstrated nuclear labeling for TFE3 protein by immunohistochemistry. The translocation was confirmed in 2 of these 5 patients by conventional cytogenetics. One case was a high-grade nonpapillary RCC and the other was compatible with type 2 papillary RCC. Four patients showed at least 1 chromosomal gain including trisomy 7 and/or trisomy 17. None of the tumors from male patients showed evidence of loss of the Y chromosome, but 2 patients showed numerical abnormalities of X chromosome +add(X). Two patients had sickle cell disease, and 1 of these also had stage IV-S neuroblastoma. This study suggests that many cases of RCC in children reported under the terms "papillary" and "clear cell" likely represent Xp11.2 translocation/TFE3 gene fusion-associated RCC. It also emphasizes the unusual associations of RCC with neuroblastoma and sickle cell hemoglobinopathy, which need further study.

Palliative Radiotherapy in Children with Neuroblastoma

Limited information is available regarding the efficacy of external beam radiation therapy in the palliation of metastatic disease from neuroblastoma. From 1960 to 2000, 29 children with 53 metastatic sites received palliative radiotherapy. There were 26 soft tissue (group I), 19 bone (group II), 5 brain (group III), and 3 hepatic (group IV) treated sites. Median radiotherapy doses for groups I, II, III, and IV sites were 2000, 2000, 2400, and 450 cGy, respectively. For group I sites, complete response was complete disappearance of mass, partial response was S 50% resolution of mass, no response was <50% resolution or h 25% progression of mass, and progressive disease was >25% progression of mass. For group II sites, complete response was complete pain relief without medication, partial response was S 50% pain relief with or without medication, no response was <50% change in pain with medication, and progressive disease was increase in pain and/or medication. Median survival was 2.5 months after palliative radiotherapy. For group I sites, complete response was seen in 1 (4%) while partial response was documented in 19 (73%). Duration of response was until death in 18 responders (90%); 2 patients relapsed with an increasing soft tissue mass at 5 months and 1 year after palliative radiotherapy. For group II sites, complete response was seen in 8 (42%) while partial response was documented in 7 (37%). Duration of response was until death in 14 responders (93%); 1 patient had relapse of pain 1 year after palliative radiotherapy. For the 5 group III children, the median survival was 2.5 months with a range of 2 days to 13 months. Four children had neurological improvement after cranial radiotherapy; one patient progressed and died secondary to neurological compromise 2 days after radiotherapy. For the 3 group IV sites, 2 had improvement in respiratory status after radiotherapy, whereas 1 progressed despite doxorubicin, cyclophosphamide, and radiation. The only patient who survived had a stage IV-S neuroblastoma with liver metastases and is alive 13 years after hepatic irradiation. Radiotherapy is an effective treatment for palliation of symptomatic metastatic disease in children with neuroblastoma.

Spontaneous Progression of a Stage IV-S Human Neuroblastoma Cell Line Involves the Increased Expression of the Protooncogenes RAS and Eukaryotic Initiation Factor 4E

Until now, there have been no established cell lines of stage IV-S neuroblastoma, a stage of neuroblastoma that can spontaneously regress. We report a newly established and characterized stage IV-S human neuroblastoma cell line (NB-UK-LO, or LO). After four initial passages of this cell line in culture, freezing, then six more passages, spontaneous transformation to a very aggressive phenotype (NB-UK-LP, or LP) was noted with evident alterations in both in vivo and in vitro characteristics. Interestingly, the untreated IV-S tumor in the patient from which the cell line was derived subsequently progressed to a more aggressive (stage IV) neuroblastoma, but it responded favorably to a short course of chemotherapy. In our report, the two cell lines are characterized in regard to in vivo growth in nude mice and molecular expression of MMP-2, MMP-9, N-ras, nm23-Hl, eIF-4E, and CD44.

Prenatally detected tumor mass in the adrenal gland

Background/Purpose : Screening programs using urinary vanillylmandelic acid have detected neuroblastomas in early infancy with some success. With the widespread use of ultrasonography in modern obstetric practice, use of ultrasonography to screen for fetal neuroblastoma seems to be reasonable and practical. Methods : Seven fetuses had suprarenal masses detected by maternal ultrasound scan at 32 to 37 weeks' gestation between 1993 and 1998. They were delivered normally if the pregnancy was uncomplicated, especially if it was without maternal preeclampsia or fetal hydrops. Each mass was further confirmed by ultrasound scan, computed tomography, or magnetic resonance imaging in the neonatal period. Tumor excision was performed at the age of 6 to 38 days of life. Results : The size of the masses measured ranged from 2.0 × 2.0 cm to 4.5 × 4.5 cm. The diagnosis was adrenal hemorrhage in 1 neonate, Evan's stage I neuroblastoma in 3, and stage IV-S neuroblastoma in 3. All of the specimens with a diagnosis of neuroblastoma showed a favorable histology by the Shimada classification system. Infants with stage I disease were treated with tumor excision only, and they had survived without disease by 14, 18, and 25 months of follow-up. One infant with stage IV-S neuroblastoma was treated further with minimal chemotherapy and has survived without disease at the 66-month follow-up examination. Another child with stage IV-S neuroblastoma has survived with local recurrence and increasing liver metastasis and was still on chemotherapy at the 2-month follow-up examination. The third child with stage IV-S disease presented with massive hepatomegaly and bone marrow involvement, and disseminated intravascular coagulopathy had developed. The patient died on the 5th day of life without surgical intervention. Conclusions : The increasing use of obstetric ultrasonography has made the prenatal screening of neuroblastomas possible. The prognosis of infants with a suprarenal mass may be improved with this early detection and early surgical intervention.

Neonatal oncology: Diagnostic and therapeutic dilemmas

Tumors are a rare but important cause of mortality and morbidity in the neonate. There is a wide spectrum of benign and malignant tumors that can occur, some of which have a unique presentation and behavior in this age group. It is important to recognize that tumors such as stage IVS neuroblastoma, which have a good prognosis in older children, may be rapidly fatal in the newborn and constitute an oncological emergency. Other tumors behave in a less aggressive fashion at this age than in the older child, and should not be overtreated. An increasing number of tumors are being detected by prenatal screening, and understanding of their natural history and management is important. Management of neonatal cancer requires knowledge also of the developmental biology of the neonate, of the way in which they handle drugs, and the increased potential for acute toxicity and long-term morbidity from chemotherapy and radiation therapy.

Stage IVS neuroblastoma: A clinical, histological, and biological analysis of 45 cases

This study reports clinicopathologic and biological analysis of 45 stage IVS neuroblastomas (IVS-NBs). All IVS-NB cases had small or undetectable primary tumors associated with disease involving the liver, skin, or bone marrow. Their ages at diagnosis ranged from 1 day to 12 months, with a median age of 4 months. The primary sites were adrenal in 34 cases, retroperitoneum in two, mediastinum in one, and unknown in eight; the liver was involved in 24 of 31 cases, the bone marrow in 11 of 27, and the skin in 4 of 29. According to the histopathological evaluation (Shimada classification), 43 IVS-NB tumors had a favorable histology, and 2 cases showed an unfavorable histology. Three patients died of progressive disease and six of indirect tumor death. Among these three deceased cases with tumor progression, two cases showed unfavorable histology, and one case had amplified N-myc (20 copies). These data suggest that most IVS-NB can be described as a tumor presenting characteristic dissemination pattern in the younger age group, having a favorable histology and showing an excellent outcome of the patients. However, poor prognosis group in stage IVS does exist, and this group could be detected histopathologically and biologically.

Resection of primary tumor at diagnosis in stage IV-S neuroblastoma: does it affect the clinical course?

To determine whether resection of primary tumor has a favorable influence on outcome of infants (age 0 to 11 months) with stage IV-S neuroblastoma. Between March 1976 and December 1993, 97 infants with previously untreated neuroblastoma diagnosed in 21 Italian institutions were classified as having stage IV-S disease. Seventy percent were younger than 4 months. Adrenal was the primary tumor site in 64 of 85 patients with a recognizable primary tumor. Liver was the organ most often infiltrated by the tumor (82 patients), followed by bone marrow and skin. The overall survival (OS) rate at 5 years in 80% and event-free survival (EFS) rate 68%. In 24 infants, the effect of resection of primary tumor could not be evaluated because of rapidly fatal disease progression (n = 8), absence of a primary tumor (n = 12), or partial resection (n = 4). Of 73 assessable patients, 26 underwent primary tumor resection at diagnosis: one died of surgical complications, one relapsed locally and died, and two others relapsed (one of these two locally) and survived, for a 5-year OS rate of 92% and EFS rate of 84%. Of the remaining 47 patients who did not undergo primary tumor resection at diagnosis 11 suffered unfavorable events, of whom five died, for an OS rate of 89% and EFS rate of 75% (no significant difference from previous group). Disease recurred at the primary tumor site in only one five who died, and in only one of six survivors of progression or relapse; in these patients, the primary tumor, located in the mediastinum, was successfully resected. Infants who underwent resection of the primary tumor at diagnosis had no better outcome than those in whom the decision was made not to operate.

Molecular characterization of a (1;10)(p22;q21) constitutional translocation from a patient with neuroblastoma

A constitutional t(1;10)(p22;q21) from a patient with stage IVS neuroblastoma has been isolated in somatic cell hybrids and the position of the breakpoints analyzed. On chromosome 1 the breakpoint lies in a 4-Mbp region flanked by minisatellite marker D1S234 and by D1S188, which lies in the region of F3. The chromosome 10 breakpoint lies between RBP3 and NAKNR below the MEN2 locus. Because, in patients with genetic disease, constitutional translocations frequently interrupt genes which are related to the clinical phenotype this rearrangement has identified two regions which potentially contain genes related to the development of neuroblastoma.

Neuroblastoma IV‐S Followed by Extra‐Adrenal Pheochromocytoma 15 Years Later

Common origin of sympathoblasts and pheochromoblasts from the neural crest cells is generally accepted. Neuroblastoma and pheochromocytoma result from an abnormal proliferation of these committed cells. They are included in the group of neuroendocrine neoplasms, formerly named Apudomas. Previous reports of mixed tumours of neuroblastoma and pheochromocytoma, and ganglioneuroma and pheochromocytoma, support this hypothesis. A case of extra-adrenal pheochromocytoma in an adolescent who 15 years before was successfully treated for a stage IV-S neuroblastoma without evidence of primitive tumour is reported. Two hypothesis are formulated: (1) the second neoplasm may have arisen from the unlocated primitive tumour, probably silent evolution could be due to its maturation into ganglioneuroma-pheochromocytoma; (2) taken into account a common origin of both neoplasms, the patient may have a genetic predisposition to develop neuroendocrine tumours. Whatever the mechanism, second neoplasms, as the case reported, late relapses, and late toxic effects may justify long follow-up of neuroblastoma survivors.

High nm23‐h1/ndpk‐a expression in ewing tumors: Paradoxical immunohistochemical reactivity and lack of prognostic significance

Expression of nm23-H1/NDPK-A has been reported to correlate inversely with metastasizing potential of rodent experimental cells and some human tumors. In the search for reliable molecular prognostic indicators for Ewing tumors (ET), a group of aggressive presumably neuroectodermal malignancies in children and adolescents, we studied nm23-H1/NDPK-A expression. Northern-blot and RT-PCR analyses were employed to semi-quantificatively measure nm23-H1 mRNA levels in ET cell lines and tissue extracts. A panel of monoclonal antibodies (MAbs) were used to evaluate protein abundance by Western blotting and immunohistochemistry. The nm23-H1/NDPK-A gene was also investigated on the DNA level to define possible genomic alterations. Our results revealed neither nm23-H1 allelic loss nor gene amplification and failed to show any significant variation in nm23-H1 mRNA or NDPK-A protein levels of primary or metastatic ET. NDPK-A protein levels were high and comparable to those of MCF-7 breast-cancer cells and to aggressive stage-IV neuroblastoma cell lines. nm23-H2/NDPK-B expression in ET was slightly more variable but generally lower than in MCF-7 cells. In the immunohistochemical analysis, however, discrepancies in the reactivity patterns with different antibodies were observed. Differential sensitivity to various fixation methods and heat treatment pointed to a structurally polymorphic NDPK-A protein. nm23-H1 expression studies using immunohistochemistry for prognostic counselling should thus be interpreted with caution.

Abdominal expansion as a bridging technique in stage IV-S neuroblastoma with massive hepatomegaly

Stage IV-S neuroblastoma has a relatively favorable outcome. However, urgent surgical treatment may be necessary for management of life-threatening complications related to massive hepatomegaly caused by metastatic tumor infiltration. The enlarged liver often becomes of primary concern because diaphragmatic elevation results in life-threatening respiratory embarrassment. An external SILASTIC ® dome has been used as a temporizing procedure to decrease the intraabdominal pressure. The authors developed a modification of this approach using an internal polytetrafluoroethylene patch to create a ventral hernia. This technique decreases the potential complications of prosthetic material use, namely, risk of infection. The graft may be left in place for an extended period and removed in staged operations as the bulk of the metastatic tumor regresses.