RESUMO
PURPOSE: Although various artificial tracheas have been developed, none have proven satisfactory for clinical use. In-body tissue architecture (IBTA) has enabled us to produce collagenous tissues with a wide range of shapes and sizes to meet the needs of individual recipients. In the present study, we investigated the long-term outcomes of patch tracheoplasty using an IBTA-induced collagenous tissue membrane ("biosheet") in a beagle model. METHODS: Nine adult female beagles were used. Biosheets were prepared by embedding cylindrical molds assembled with a silicone rod and a slitting pipe into dorsal subcutaneous pouches for 2 months. The sheets were then implanted by patch tracheoplasty. An endoscopic evaluation was performed after 1, 3, or 12 months. The implanted biosheets were harvested for a histological evaluation at the same time points. RESULTS: All animals survived the study. At 1 month after tracheoplasty, the anastomotic parts and internal surface of the biosheets were smooth with ciliated columnar epithelium, which regenerated into the internal surface of the biosheet. The chronological spread of chondrocytes into the biosheet was observed at 3 and 12 months. CONCLUSIONS: Biosheets showed excellent performance as a scaffold for trachea regeneration with complete luminal epithelium and partial chondrocytes in a 1-year beagle implantation model of patch tracheoplasty.
Assuntos
Materiais Biocompatíveis , Membranas Artificiais , Procedimentos de Cirurgia Plástica/métodos , Engenharia Tecidual , Traqueia/cirurgia , Estenose Traqueal/cirurgia , Animais , Modelos Animais de Doenças , Cães , Feminino , Fatores de Tempo , Resultado do TratamentoRESUMO
BACKGROUND: The aim of the present study was to compare the efficacy, complications and outcomes of the following two surgical strategies for delayed local treatment for International Neuroblastoma Staging System (INSS) 4 neuroblastoma (NB): complete resection (CR; period A); and gross total resection/subtotal resection (GTR/STR) with local irradiation (period B). METHODS: We retrospectively analyzed 17 patients with INSS 4 NB who received delayed local treatment (period A, n = 11; period B, n = 6). RESULTS: Eleven patients in period A received CR. Two patients underwent GTR and four patients underwent STR in period B. The amount of blood loss in period A was significantly greater than that in period B. Postoperative complications were observed in eight patients in period A (73%), but in only one patient in period B (17%; P < 0.01). Recurrence was observed in five patients in period A and in one patient in period B (45.4% vs 16.6%; P = n.s.). Distant metastasis at recurrence was observed in four patients in period A and in one patient in period B. CONCLUSIONS: Gross total resection/subtotal resection with local irradiation may be a safe and effective delayed local treatment for INSS 4 NB.
Assuntos
Neoplasias das Glândulas Suprarrenais/cirurgia , Neuroblastoma/cirurgia , Neoplasias Retroperitoneais/cirurgia , Neoplasias das Glândulas Suprarrenais/patologia , Neoplasias das Glândulas Suprarrenais/radioterapia , Criança , Pré-Escolar , Feminino , Seguimentos , Humanos , Lactente , Masculino , Estadiamento de Neoplasias , Neuroblastoma/patologia , Neuroblastoma/radioterapia , Radioterapia Adjuvante , Neoplasias Retroperitoneais/patologia , Neoplasias Retroperitoneais/radioterapia , Estudos Retrospectivos , Resultado do TratamentoRESUMO
We report a case of successful magnetic compression anastomosis (MCA) for obstructed cyst-jejunostomy in a young woman who had undergone surgery for type 1 biliary atresia (BA) on day 78 of life. A 16-year-old girl was admitted with obstructive jaundice. Jaundice resolved with percutaneous trans-hepatic cholangiodrainage (PTCD) but contrast medium injected from the PTCD tube did not flow through the anastomosis. Magnets were placed on each side of the anastomosis, in the cyst and the jejunum, to compress the partition. On postoperative day (POD) 6, the anastomosis was recanalized and the PTCD tube placed trans-anastomotically until POD 245. The patient remained free from jaundice after removal of the PTCD tube. MCA can be a useful and less invasive procedure for treating biliary tract anastomotic obstruction in patients with BA.
Assuntos
Atresia Biliar/cirurgia , Obstrução Intestinal/cirurgia , Icterícia Obstrutiva/cirurgia , Doenças do Jejuno/cirurgia , Imãs , Complicações Pós-Operatórias/cirurgia , Adolescente , Anastomose Cirúrgica , Drenagem/métodos , Feminino , Humanos , Obstrução Intestinal/etiologia , Icterícia Obstrutiva/etiologia , Doenças do Jejuno/etiologia , JejunostomiaRESUMO
We herein report the findings of 3 patients with primary Ewing sarcoma in a pelvic lesion who underwent the placement of a tissue expander (TE) before radiation therapy to prevent the adverse effects of radiotherapy. The simulation study showed that the TE drastically reduced volume of the intestine that was irradiated at all dose levels. All patients could receive the scheduled dose of radiotherapy without any acute and late complications such as diarrhea, melena, the dislodging of the TE, infection, or the formation of fistulae. In the 4-year (minimum) observation period, we did not observe intestinal complications in any of our patients. TE placement is considered to be a safe and effective method for preventing the adverse effects of radiotherapy in pediatric malignant pelvic tumors.
Assuntos
Neoplasias Ósseas/radioterapia , Intestinos/efeitos da radiação , Neoplasias Pélvicas/radioterapia , Lesões por Radiação/prevenção & controle , Sarcoma de Ewing/radioterapia , Dispositivos para Expansão de Tecidos , Adolescente , Criança , Feminino , Humanos , Radioterapia/efeitos adversosRESUMO
BACKGROUND: We have introduced and performed percutaneous sclerotherapy on pediatric patients, and information regarding the mid- and long-term results after percutaneous treatment of peripheral venous malformations is necessary to counsel patients and their parents about the outcome of the therapy. This study was designed to retrospectively evaluate the long-term satisfaction of pediatric patients following percutaneous sclerotherapy for venous malformations (VMs). METHODS: A total of 53 children younger than 16 years of age with VMs who underwent sclerotherapy were included in this study. Self-assessment questionnaires regarding the treatment effectiveness and patients' satisfaction were sent to 50 of 53 patients. Sclerotherapy was performed using 3% polidocanol, absolute ethanol, or 5% ethanolamine oleate (EO). RESULTS: The median numbers of sessions per patient was 2.0 (range, 1-12), and the median follow-up time was 1.8 years (range, 6months-8.3years). Forty-two patients (84%) returned the questionnaire. The most frequent symptom was swelling (95%, 40/42) before sclerotherapy. After treatment, patients felt less pain (82%, 23/28), functional limitation (78%, 14/18), swelling (70%, 28/40), and cosmetic disfigurement (59%, 13/22). None of the patients responded "worse" for any symptoms, and 30 patients (71%) reported being "very satisfied" or "satisfied" with the treatment. CONCLUSION: Percutaneous sclerotherapy of VMs was safe and effective for relieving symptoms in our pediatric patients, and many of them were satisfied with the outcomes.
Assuntos
Satisfação do Paciente , Polietilenoglicóis/administração & dosagem , Escleroterapia/métodos , Malformações Vasculares/tratamento farmacológico , Veias/anormalidades , Adolescente , Criança , Pré-Escolar , Feminino , Seguimentos , Humanos , Injeções Intralesionais , Imageamento por Ressonância Magnética , Masculino , Flebografia , Polidocanol , Estudos Retrospectivos , Soluções Esclerosantes/administração & dosagem , Inquéritos e Questionários , Fatores de Tempo , Resultado do Tratamento , Malformações Vasculares/diagnóstico , Veias/diagnóstico por imagemRESUMO
In pediatric LDLT, graft reduction is sometimes required because of the graft size mismatch. Dividing the portal triad and hepatic veins with a linear stapler is a rapid and safe method of reduction. We herein present a case with a left lateral segment reduction achieved using a linear stapler after reperfusion in pediatric LDLT. The patient was a male who had previously undergone Kasai procedure for biliary atresia. We performed the LDLT with his father's lateral segment. According to the pre-operative volumetry, the GV/SLV ratio was 102.5%. As the patient's PV was narrow, sclerotic and thick, we decided to put an interposition with the IMV graft of the donor between the confluence and the graft PV. The graft PV was anastomosed to the IMV graft. The warm ischemic time was 34 min, and the cold ischemic time was 82 min. The ratio of the graft size to the recipient weight (G/R ratio) was 4.2%. After reperfusion, we found that the graft had poor perfusion and decided to reduce the graft size. We noted good perfusion in the residual area after the lateral edge was clamped with an intestinal clamp. The liver tissue was sufficiently fractured with an intestinal clamp and then was divided with a linear stapler. The final G/R ratio was 3.6%. The total length of the operation was 12 h and 20 min. The amount of blood lost was 430 mL. No surgical complications, including post-operative hemorrhage and bile leakage, were encountered. We believe that using the linear stapler decreased the duration of the operation and was an acceptable technique for reducing the graft after reperfusion.
Assuntos
Atresia Biliar/cirurgia , Hepatectomia/métodos , Transplante de Fígado , Doadores Vivos , Coleta de Tecidos e Órgãos/métodos , Humanos , Lactente , Masculino , Tamanho do Órgão , Grampeamento CirúrgicoRESUMO
BACKGROUND: Parenteral nutrition (PN)-associated liver dysfunction (PNALD) in term infants usually manifests as intrahepatic cholestasis, which recovers with enteral nutrition (EN) in most cases; however, as the number of extremely low-birthweight infants (ELBWI) has been increasing, and consequently intestinal diseases associated with ELBWI have been increasing, more intractable PNALD has been encountered after surgical treatment in ELBWI, which does not resolve or rather worsens with EN. METHODS: Three cases of ELBWI with intestinal perforation, which developed PNALD and eventually died of hepatic failure with intractable portal hypertension, were reviewed. Their gestational age and birthweight ranged from 23 to 26 weeks, and from 434 to 968 g, respectively. The intestinal diseases included necrotizing enteritis in two and meconium-related ileus with focal intestinal perforation in one. RESULTS: The duration of total PN without EN in the three cases was 17, 24 and 24 days, respectively. The interval between the introduction of PN and the onset of PNALD was 14, 4 and 18 days, respectively. A marked elevation of serum endotoxin level was detected in both cases of necrotizing enteritis. Histopathological study of the liver revealed marked cholestasis, significant hepatic necrosis with fibrosis, and proliferation of ductules in all these cases, which was responsible for portal hypertension. CONCLUSIONS: PN after gastrointestinal disorders in ELBWI may cause refractory PNALD, which does not resolve, or rather worsens with the resumption of EN. Portal hypertension secondary to hepatic necrosis may be responsible for the exacerbation with the resumption of EN.
Assuntos
Nutrição Enteral/efeitos adversos , Hipertensão Portal/etiologia , Doenças do Prematuro/terapia , Perfuração Intestinal/terapia , Falência Hepática/etiologia , Nutrição Parenteral/efeitos adversos , Enterocolite Necrosante/terapia , Evolução Fatal , Humanos , Hipertensão Portal/diagnóstico , Doenças do Íleo/terapia , Íleus/terapia , Recém-Nascido de Peso Extremamente Baixo ao Nascer , Recém-Nascido , Recém-Nascido Prematuro , Doenças do Prematuro/diagnóstico , Doenças do Prematuro/etiologia , Falência Hepática/diagnóstico , MasculinoRESUMO
PURPOSE: To clarify the significance of intrahepatic cystic lesions (ICLs) after Kasai procedure for biliary atresia (BA), we reviewed the BA patients with special reference to the shape of ICLs. METHODS: For the last two decades, 75 cases have been followed up at our institution. The patients were divided into two groups: ICLs (+) with ICL and ICLs (-) without ICL. Seventeen cases of ICLs (+) were divided into two categories: the round type and the multiple-bead type. A poor prognosis was defined as mortality or the need for liver transplantation (LT). RESULTS: The ratio of round to multiple-bead types was 6:11. The percentage of poor prognoses with ICLs (+) and ICLs (-) was 47 and 53 %, respectively. The percentage of poor prognoses with round and multiple-bead types was 27 and 83 %, respectively (p < 0.05). In cases of ICLs (+), an LT was required because of hepatic deterioration in all three of the round types, and intractable cholangitis in all five of the multiple-bead types (p < 0.05). CONCLUSION: The long-term prognosis of BAs after the Kasai procedure does not necessarily depend on the development of ICLs per se, but on their shape.
Assuntos
Atresia Biliar/cirurgia , Cistos/diagnóstico por imagem , Hepatopatias/diagnóstico por imagem , Portoenterostomia Hepática/efeitos adversos , Cistos/epidemiologia , Cistos/etiologia , Seguimentos , Humanos , Incidência , Lactente , Recém-Nascido , Japão/epidemiologia , Hepatopatias/epidemiologia , Hepatopatias/etiologia , Transplante de Fígado , Complicações Pós-Operatórias , Prognóstico , Estudos Retrospectivos , Índice de Gravidade de Doença , UltrassonografiaRESUMO
Osteosarcoma, the most common pediatric bone tumor, is an aggressive heterogeneous malignancy defined by complex chromosomal aberrations. Overall survival rates remain at ~70%, but patients with chemoresistant or metastatic disease have extremely poor outcomes of <30%. A subgroup of tumors harbor amplification of chromosome 8q24.2 and increased expression of the oncogenic long noncoding RNA (lncRNA) Plasmacytoma Variant Translocation-1 (PVT-1), which is associated with an extremely poor clinical prognosis. This study demonstrates that PVT-1 is critical for osteosarcoma tumor-initiation potential. Chromatin Hybridization by RNA Purification analysis identified Tripartite-Motif Containing Family 28 (TRIM28) as a novel PVT-1 binding partner. Mechanistically, co-immunoprecipitation studies showed the PVT-1/TRIM28 complex binds and increases SUMOylation of phosphatidylinositol 3-kinase catalytic subunit type 3 (Vps34), which leads to enhanced ubiquitination and degradation of tumor suppressor complex 2 (TSC2), thus contributing to increased self-renewal and stem cell phenotypes. Furthermore, we identified that osteosarcoma cells with increased PVT-1 have enhanced sensitivity to the SUMOylation inhibitor, TAK-981. Altogether, this study elucidated a role for PVT-1 in the enhancement of cancer stem-like behaviors, including migration and invasion, in osteosarcoma, and identified the novel PVT-1/TRIM28 axis signaling cascade as a potential therapeutic target for osteosarcoma treatment.
Assuntos
Neoplasias Ósseas , Osteossarcoma , RNA Longo não Codificante , Proteína 28 com Motivo Tripartido , Proteína 2 do Complexo Esclerose Tuberosa , Humanos , Neoplasias Ósseas/patologia , Linhagem Celular Tumoral , Proliferação de Células/genética , Regulação Neoplásica da Expressão Gênica , Osteossarcoma/patologia , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Transdução de Sinais/genética , Proteína 28 com Motivo Tripartido/genética , Proteína 2 do Complexo Esclerose Tuberosa/metabolismoRESUMO
Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma in children, with overall long-term survival rates of â¼65-70%. Thus, additional molecular insights and representative models are critical for identifying and evaluating new treatment modalities. Using MyoD-Cre-mediated introduction of mutant K-RasG12D and perturbations in p53, we developed a novel genetically engineered mouse model (GEMM) for RMS. The anatomic sites of primary RMS development recapitulated human disease, including tumors in the head, neck, extremities and abdomen. We confirmed RMS histology and diagnosis through Hematoxylin and Eosin staining, and positive immunohistochemical staining for desmin, myogenin, and phosphotungstic acid-Hematoxylin. Cell lines from GEMM tumors were established with the ability to engraft in immunocompetent mice with comparable histological and staining features as the primary tumors. Tail vein injection of cell lines had high metastatic potential to the lungs. Transcriptomic analyses of p53R172H/K-RasG12D GEMM-derived tumors showed evidence of high molecular homology with human RMS. Finally, pre-clinical use of these murine RMS lines showed similar therapeutic responsiveness to chemotherapy and targeted therapies as human RMS cell lines.
Assuntos
Rabdomiossarcoma , Sarcoma , Neoplasias de Tecidos Moles , Animais , Modelos Animais de Doenças , Humanos , Camundongos , Rabdomiossarcoma/diagnóstico , Rabdomiossarcoma/tratamento farmacológico , Rabdomiossarcoma/genética , Sarcoma/metabolismo , Neoplasias de Tecidos Moles/metabolismo , Proteína Supressora de Tumor p53/genéticaRESUMO
Ewing sarcoma (ES) is the second most common bone tumor in children and young adults. Unfortunately, there have been minimal recent advancements in improving patient outcomes, especially in metastatic and recurrent diseases. In this study, we investigated the biological role of p21-activated kinases (PAKs) in ES, and the ability to therapeutically target them in high-risk disease. Via informatics analysis, we established the inverse association of PAK1 and PAK4 expression with clinical stage and outcome in ES patients. Through expression knockdown and small-molecule inhibition of PAKs, utilizing FRAX-597, KPT-9274, and PF-3758309 in multiple ES cell lines and patient-derived xenograft models, we further explored the role of PAKs in ES tumor growth and metastatic capabilities. In vitro studies in several ES cell lines indicated that diminishing PAK1 and PAK4 expression reduces tumor cell viability, migratory, and invasive properties. In vivo studies using PAK4 inhibitors, KPT-9274 and PF-3758309 demonstrated significant inhibition of primary and metastatic tumor formation, while transcriptomic analysis of PAK4-inhibitor-treated tumors identified concomitant suppression of Notch, ß-catenin, and hypoxia-mediated signatures. In addition, the analysis showed enrichment of anti-tumor immune regulatory mechanisms, including interferon (IFN)-É£ and IFN-α responses. Altogether, our molecular and pre-clinical studies are the first to establish a critical role for PAKs in ES development and progression, and consequently as viable therapeutic targets for the treatment of high-risk ES in the near future.
Assuntos
Sarcoma de Ewing/tratamento farmacológico , Quinases Ativadas por p21/genética , Acrilamidas/farmacologia , Aminopiridinas/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Interferon-alfa/genética , Interferon gama/genética , Pirazóis/farmacologia , Pirróis/farmacologia , Sarcoma de Ewing/genética , Sarcoma de Ewing/patologia , Transdução de Sinais/efeitos dos fármacos , Ensaios Antitumorais Modelo de Xenoenxerto , Quinases Ativadas por p21/antagonistas & inibidoresRESUMO
The use of a bioartificial liver with pig liver cells in the treatment of fulminant hepatic failure has already begun as a clinical trial in several countries. Therefore, studies on plasma complement regulatory proteins of the pig are necessary, because the liver produces them. Complement factor I is a serine protease that cleaves C3b and C4b. The DNA sequences of factor I have been reported in many species, with the noted exception of pigs. In this study, porcine factor I was cloned and an open reading frame of 1794 base pairs were identified. The predicted amino acid sequence maintained a relatively high homology compared to those of other mammals, especially in the serine protease (SP) region. The cell membrane-bound forms of the porcine and the human SP domain of factor I were constructed. Amelioration of complement-mediated cell lysis by these molecules was then tested, using several kinds of sera and Chinese hamster ovary (CHO) cell transfectants. Both molecules had a suppressing effect on pig, human and dog complements, indicating little species-specificity. Further studies of other plasma complement regulatory proteins produced from pig liver cells will need to be considered as the primary feature of the pig bioartificial liver.
Assuntos
Fator I do Complemento/imunologia , Fator I do Complemento/metabolismo , Hepatócitos/imunologia , Suínos/imunologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Células CHO , Clonagem Molecular , Fator I do Complemento/química , Fator I do Complemento/genética , Proteínas do Sistema Complemento/imunologia , Proteínas do Sistema Complemento/metabolismo , Cricetinae , Cricetulus , Cães , Hepatócitos/metabolismo , Humanos , Dados de Sequência Molecular , Alinhamento de Sequência , Serina Endopeptidases/imunologia , Serina Endopeptidases/metabolismo , Especificidade da Espécie , Transplante HeterólogoRESUMO
OBJECTIVE: To present the efficacy of Japanese-traditional medicine (Kampo) for a case with vascular malformation. METHODS: A case study and literature review. PATIENT: A 62-year-old female presented with dysphagia and spitting blood. Esophagogastroduodenoscopy showed a longitudinal lobulated and septated mass in the posterior pharynx. On MR imaging, the mass showed hyperintensity on T2-weighted images and heterogeneous enhancement on Gadlinium-enhanced T1-weighted images, suggestive of a low-flow vascular malformation. INTERVENTION: According to the Kampo diagnosis, kamisyouyousan and ninjinyoueito were prescribed to this patient. The effect of Kampo medicine was evaluated with improvement of her symptoms and volumetry of MRI findings. RESULT: The longitudinal pharyngeal mass was markedly decreased and her symptoms disappeared after 2 years of Kampo administration. CONCLUSIONS: Kampo medicine can be a novel alternative therapy for VM.
Assuntos
Medicamentos de Ervas Chinesas/uso terapêutico , Medicina Kampo , Faringe/irrigação sanguínea , Malformações Vasculares/tratamento farmacológico , Feminino , Hemoptise/etiologia , Humanos , Angiografia por Ressonância Magnética , Pessoa de Meia-Idade , Faringe/diagnóstico por imagem , Fitoterapia , Malformações Vasculares/complicações , Malformações Vasculares/diagnóstico por imagemRESUMO
PURPOSE: Cisplatin is an effective chemotherapeutic agent used to treat a variety of malignant tumors. The major toxicity associated with cisplatin treatment is granulocytopenia. C-type natriuretic peptide (CNP), a member of the natriuretic peptide family, protects against toxicity in many organs, including the heart, blood vessels, lung, and kidney. The objective of this study was to investigate the myeloprotective effects of CNP in a mouse model of cisplatin-induced granulocytopenia. METHODS: The mice were divided into two groups: cisplatin with vehicle and cisplatin with CNP. CNP (2.5 µg/kg/min via osmotic pump, subcutaneously) or vehicle administration was started two day before cisplatin injection, and continued until the mice were killed. At 0, 2, 4, 8, and 14 days after cisplatin injection (16 mg/kg, intraperitoneally as a single dose), we counted total and living cells and granulocyte/macrophage colony-forming units (CFU-GM) in bone marrow. In addition, at 0, 1, 2, and 4 days after cisplatin injection, we measured mRNA levels of CXC chemokine receptor 4 (CXCR4) and chemokine CXC ligand 12 (CXCL12) in bone marrow. RESULTS: CNP significantly attenuated the reduction in bone marrow nucleated cell count and CFU-GM in bone marrow at 4 days after cisplatin injection. Four days after cisplatin injection, CNP significantly decreased the CXCR4 mRNA level in bone marrow, but had no effect on the level of CXCL12 mRNA. CONCLUSIONS: CNP exerts myeloprotective effects in cisplatin-induced granulocytopenia and decreases CXCR4 expression.
Assuntos
Agranulocitose/prevenção & controle , Medula Óssea/efeitos dos fármacos , Cisplatino/toxicidade , Peptídeo Natriurético Tipo C/farmacologia , Animais , Quimiocina CXCL12/genética , Camundongos , Camundongos Endogâmicos C57BL , Substâncias Protetoras/farmacologia , RNA Mensageiro/análise , Receptores CXCR4/genéticaRESUMO
The pig cDNA encoding C1 esterase inhibitor (C1-INH) was isolated and the homology of the sequence was compared with that from other animals. The structure of pig C1-INH contains a two disulfide bridge pattern identical to the human C1-INH. In the amino acid sequence of the first Cys-91 to the C-terminal end, the pigC1-INH has a 76.2% homology with the human protein, and the sequence of the reactive site is close to the human. A surface-bound form of pig and human C1-INH, pC1-INH-PI and hC1-INH, respectively, were next constructed. Stable Chinese hamster ovarian tumor (CHO) cell lines and pig endothelial cell (PEC) lines expressing these C1-INH-PI were prepared by transfection. The basic function and the species specificity of pCI-INH were then investigated using these transfectants. pC1-INH and hC1-INH have almost the same suppressive effect on pig, human, dog and rabbit sera in complement-dependent cell lysis, indicating little species specificity.
Assuntos
Proteínas Inativadoras do Complemento 1/genética , Sus scrofa/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Células CHO , Clonagem Molecular , Cricetinae , Cricetulus , Primers do DNA , DNA Complementar/genética , Citometria de Fluxo , Humanos , Immunoblotting , Dados de Sequência Molecular , Alinhamento de Sequência , Análise de Sequência de DNA , Especificidade da EspécieRESUMO
Survivin, a member of the inhibitor of apoptosis protein family, functions as a key regulator of programmed cell death. YM155 is a small molecule that selectively inhibits survivin. We investigated the effect of YM155 on survivin suppression in the human rhabdomyosarcoma (RMS) cell line RD. The efficacy of YM155 in combination with cisplatin was also determined in a xenograft model. The effect of YM155 on survivin expression in the RD cell line was examined at both mRNA and protein levels using real-time PCR and western blot analysis. RD cells were cultured with various concentrations of YM155, then cisplatin was added to the medium and the anti-proliferation response was determined. Cell growth was evaluated by WST-8 assay. Finally, the efficacy of YM155 combined with cisplatin was examined in an established xenograft model. Survivin mRNA levels in the RD cell line were decreased to 72 and 24% at 24 and 48 h, respectively, after 10 nM of YM155 was added. YM155 also decreased the levels of survivin protein. YM155 treatment (10 nM) inhibited cell proliferation of RD in a dose-dependent manner in vitro, with 58% of cells viable at 48 h. When cultured with 10 nM of YM155 and 10 µM cisplatin, RD cells demonstrated only 25% of the growth observed when cultured with cisplatin alone. YM155 in combination with cisplatin significantly inhibited tumor growth by 13% compared with control (P<0.0001) in RD xenograft tumors. YM155 increased the sensitivity of cisplatin by suppressing survivin in the embryonal RMS cell line RD. Further studies should investigate the use of YM155 as an apoptosis inducer, either alone or in combination with cisplatin, for the treatment of malignant RMS.
Assuntos
Cisplatino/administração & dosagem , Imidazóis/administração & dosagem , Proteínas Inibidoras de Apoptose/genética , Naftoquinonas/administração & dosagem , Rabdomiossarcoma Embrionário/tratamento farmacológico , Rabdomiossarcoma Embrionário/genética , Animais , Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Apoptose , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Cisplatino/farmacologia , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Imidazóis/farmacologia , Proteínas Inibidoras de Apoptose/metabolismo , Camundongos , Naftoquinonas/farmacologia , Rabdomiossarcoma Embrionário/metabolismo , Survivina , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
CT-guided localization followed by video-assisted thoracoscopic surgery was performed in three children with pulmonary nodules less than 5 mm in diameter. The patients' respective primary diagnoses were Wilms tumor, hepatoblastoma, and osteosarcoma of the femur. The pulmonary nodules were marked preoperatively by a percutaneously placed hook-wire or dye under CT guidance. Although none of the nodules was grossly detected during the operation, they were correctly resected under the guidance of the hook-wire wound or dye. A histological assessment revealed viable metastatic lesions in the case of hepatoblastoma, completely necrotic lesions in the case of Wilms tumor, and inflammatory nodules in the case of osteosarcoma. CT-guided localization followed by video-assisted thoracoscopic surgery appears to be a beneficial procedure in children with tiny pulmonary nodules.
Assuntos
Neoplasias Pulmonares/diagnóstico por imagem , Neoplasias Pulmonares/cirurgia , Nódulo Pulmonar Solitário/diagnóstico por imagem , Nódulo Pulmonar Solitário/cirurgia , Cirurgia Torácica Vídeoassistida , Tomografia Computadorizada por Raios X , Adolescente , Pré-Escolar , Feminino , HumanosRESUMO
Rhabdomyosarcoma (RMS) is a malignant mesenchymal tumor and the most common soft tissue sarcoma in children. Because of several complications associated with intensive multimodal therapies, including growth disturbance and secondary cancer, novel therapies with less toxicity are urgently needed. C-type natriuretic peptide (CNP), an endogenous peptide secreted by endothelial cells, exerts antiproliferative effects in multiple types of mesenchymal cells. Therefore, we investigated whether CNP attenuates proliferation of RMS cells. We examined RMS patient samples and RMS cell lines. All RMS clinical samples expressed higher levels of guanylyl cyclase B (GC-B), the specific receptor for CNP, than RMS cell lines. GC-B expression in RMS cells decreased with the number of passages in vitro. Therefore, GC-B stable expression lines were established to mimic clinical samples. CNP increased cyclic guanosine monophosphate (cGMP) levels in RMS cells in a dose-dependent manner, demonstrating the biological activity of CNP. However, because cGMP is quickly degraded by phosphodiesterases (PDEs), the selective PDE5 inhibitor sildenafil was added to inhibit its degradation. In vitro, CNP, and sildenafil synergistically inhibited proliferation of RMS cells stably expressing GC-B and decreased Raf-1, Mitogen-activated protein kinase kinase (MEK), and extracellular signal-regulated kinase (ERK) phosphorylation. These results suggested that CNP in combination with sildenafil exerts antiproliferative effects on RMS cells by inhibiting the Raf/MEK/ERK pathway. This regimen exerted synergistic effects on tumor growth inhibition without severe adverse effects in vivo such as body weight loss. Thus, CNP in combination with sildenafil represents a promising new therapeutic approach against RMS.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Rabdomiossarcoma/patologia , Adolescente , Animais , Proliferação de Células/efeitos dos fármacos , Criança , Pré-Escolar , Relação Dose-Resposta a Droga , Sinergismo Farmacológico , Feminino , Humanos , Lactente , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Masculino , Camundongos Endogâmicos BALB C , Peptídeo Natriurético Tipo C/administração & dosagem , Fosforilação/efeitos dos fármacos , Receptores do Fator Natriurético Atrial/metabolismo , Rabdomiossarcoma/enzimologia , Citrato de Sildenafila/administração & dosagem , Células Tumorais Cultivadas , Regulação para Cima , Ensaios Antitumorais Modelo de Xenoenxerto/métodosRESUMO
Cancer stem cells (CSCs) are defined as a small population of cancer cells with the properties of high self-renewal, differentiation, and tumor-initiating functions. Recent studies have demonstrated that aldehyde dehydrogenase 1 (ALDH1) is a marker for CSCs in adult cancers. Although CSCs have been identified in some different types of pediatric solid tumors, there have been no studies regarding the efficacy of ALDH1 as a marker for CSCs. Therefore, in order to elucidate whether ALDH1 can be used as a marker for CSCs of pediatric sarcoma, we examined the characteristics of a population of cells with a high ALDH1 activity (ALDH1high cells) in rhabdomyosarcoma (RMS), the most common soft tissue sarcoma in children. We used the human embryonal RMS (eRMS) cell lines RD and KYM-1, and sorted the cells into two subpopulations of ALDH1high cells and cells with a low ALDH1 activity (ALDH1low cells). Consequently, we found that the ALDH1high cells comprised 3.9% and 8.2% of the total cell population, respectively, and showed a higher capacity for self-renewal and tumor formation than the ALDH1low cells. With regard to chemoresistance, the survival rate of the ALDH1high cells was found to be higher than that of the ALDH1low cells following treatment with chemotherapeutic agents for RMS. Furthermore, the ALDH1high cells exhibited a higher degree of pluripotency and gene expression of Sox2, which is one of the stem cell markers. Taken together, the ALDH1high cells possessed characteristics of CSCs, including colony formation, chemoresistance, differentiation and tumor initiation abilities. These results suggest that ALDH1 is a potentially useful marker of CSCs in eRMS.