RESUMO
Six pigs underwent implantation of a portal vein infusion port by transjugular access. The technical success rate was 100% (n = 6), with no surgical complications or deaths. At 1 month after implantation, the catheter tip had moved from the splenic vein to the main portal vein, while the catheter protruded into the right ventricle through the right atrium in all cases. Hence, the infusion port system has not been used in clinical practice due to its obvious displacement after implantation. However, this study provides a new idea for future exploration of portal vein infusion pathways.
Assuntos
Cateterismo Periférico/instrumentação , Veias Jugulares , Veia Porta , Dispositivos de Acesso Vascular , Animais , Cateterismo Periférico/efeitos adversos , Desenho de Equipamento , Estudos de Viabilidade , Feminino , Infusões Intravenosas , Veias Jugulares/diagnóstico por imagem , Masculino , Veia Porta/diagnóstico por imagem , Punções , Sus scrofa , Fatores de TempoRESUMO
BACKGROUND: Gene therapy is regarded as a new and promising therapeutic modality for cancers, and adenovirus is one of the most frequently used vectors. However, because of low or absent coxsackievirus and adenovirus receptor levels on the surface of many kinds of tumor cells, the efficiency of adenovirus infection of target tumor cells may be low. Meanwhile, gene therapy by a single vector carrying two or more antioncogenes can improve treatment effects and reduce side effects from vectors. In this research, we aimed to detect the antitumor effect of ING4/PTEN double tumor suppressors mediated by adenovirus modified with arginine(R)-glycine(G)-aspartate(D) (RGD) on glioma cells. METHODS: We treated U87 glioma cells with PBS, blank adenovirus or adenovirus carrying RGD, ING4, PTEN, or both ING4 and PTEN, then we detected and compared the U87 cells' growth, apoptosis, and invasion. Moreover, we established a U87 glioma transplantation tumor model to study the antitumor effect in vivo by measuring the volume and weight of tumor masses in each condition. In addition, we analyzed the transcription of related genes by fluorescent quantitative PCR and detected their expression by immunohistochemistry staining to reveal the underlying mechanisms. RESULTS: The double tumor suppressors ING4/PTEN could inhibit the growth of U87 glioma cells with a synergistic antitumor effect, and the RGD modification also acted as an antioncogene to inhibit U87 cell invasion and tumor angiogenesis. CONCLUSIONS: The ING4/PTEN double tumor suppressors mediated by adenovirus modified with RGD had a significantly synergistic antitumor effect on glioma.
Assuntos
Infecções por Adenoviridae/terapia , Adenoviridae/genética , Proteínas de Ciclo Celular/antagonistas & inibidores , Glioma/terapia , Proteínas de Homeodomínio/antagonistas & inibidores , Proteínas de Membrana/antagonistas & inibidores , PTEN Fosfo-Hidrolase/antagonistas & inibidores , Proteínas Supressoras de Tumor/antagonistas & inibidores , Aminoácidos/genética , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Proliferação de Células/efeitos dos fármacos , Terapia Genética/métodos , Glioma/virologia , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Humanos , Proteínas de Membrana/genética , Neovascularização Patológica , PTEN Fosfo-Hidrolase/genética , Proteínas Supressoras de Tumor/genéticaRESUMO
Previous studies have shown that interleukin-24 (IL-24) has tumor-suppressing activity by multiple pathways. However, the immunogenicity moderation effect of IL-24 on malignant cells has not been explored extensively. In this study, we investigated the role of IL-24 in immunogenicity modulation of the myelogenous leukemia cells. Data show that myelogenous leukemia cells express low levels of immunogenicity molecules. Treatment with IL-24 could enhance leukemia cell immunogenicity, predominantly regulate leukemia cells to produce immune-associated cytokines, and improve the cytotoxic sensitivity of these cells to immune effector cells. IL-24 expression could retard transplanted leukemia cell tumor growth in vivo in athymic nude mice. Moreover, IL-24 had marked effects on downregulating the expression of angiogenesis-related proteins vascular endothelial growth factor, cluster of differentiation (CD) 31, CD34, collagen IV and metastasis-related factors CD147, membrane type-1 matrix metalloproteinase (MMP), and MMP-2 and MMP-9 in transplanted tumors. These findings indicated novel functions of this antitumor gene and characterized IL-24 as a promising agent for further clinical trial for hematologic malignancy immunotherapy.
Assuntos
Adjuvantes Imunológicos/farmacologia , Imunomodulação , Interleucinas/uso terapêutico , Leucemia Mieloide/tratamento farmacológico , Células Mieloides/efeitos dos fármacos , Inibidores da Angiogênese/imunologia , Inibidores da Angiogênese/farmacologia , Inibidores da Angiogênese/uso terapêutico , Animais , Linhagem Celular Tumoral , Células Matadoras Induzidas por Citocinas/imunologia , Células Matadoras Induzidas por Citocinas/metabolismo , Humanos , Interleucinas/imunologia , Interleucinas/farmacologia , Leucemia Mieloide/imunologia , Camundongos , Camundongos Nus , Células Mieloides/imunologia , Invasividade Neoplásica/imunologia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
To evaluate the role of microRNA-126 (miR-126) in maintaining the integrity of the blood-retina barrier (BRB), we established a mouse model of oxygen-induced retinopathy (OIR) and measured the retinal levels of miR-126 using recombinant plasmid pCMV-MIR or pCMV-MIR-126 intravitreal injections. We also detected VCAM-1 and BCL2L11 levels. Retinal vaso-obliteration, VCAM-1 localization on retinal endothelial cells, the blood-retina vascular permeability or albumin leakage in retinas, TUNEL histology, Evans blue assays, or Western blotting for detecting albumin or tight junction levels in the retina was performed. We also detected the effect of miR-126 on the survival of Müller cells in a mouse model using vimentin fluorescence staining. Our results suggested that miR-126 may not only regulate the overexpression of VCAM-1 or BCL2L11 and lead to the reduction of retinal endothelial cell apoptosis, retinal vascular leakage, or retinal permeability in the OIR mouse model, but may also protect hypoxic retinal Müller cells via the STAT3 signaling pathway. We believe that miR-126 could also be a potential therapeutic agent to maintain the stability of the BRB in ischemic retinopathy.
Assuntos
Proteína 11 Semelhante a Bcl-2/metabolismo , Barreira Hematorretiniana/fisiologia , Isquemia/metabolismo , MicroRNAs/fisiologia , Doenças Retinianas/metabolismo , Molécula 1 de Adesão de Célula Vascular/metabolismo , Animais , Animais Recém-Nascidos , Apoptose/fisiologia , Western Blotting , Permeabilidade Capilar/fisiologia , Modelos Animais de Doenças , Células Endoteliais/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Retina/citologia , Doenças Retinianas/patologia , Vasos Retinianos/metabolismo , Vasos Retinianos/patologiaRESUMO
Interleukin-24 (IL-24), a member of the IL-10 cytokine gene family, causes growth suppression and apoptosis in various solid tumor cells. However, the effects of IL-24 on hematopoietic malignant cells have not been extensively explored. In this report, we constructed an RGD-engineered recombinant adenoviral vector, Ad.RGD-IL-24, and assessed its effects on human myeloid leukemia cells. Ad vector mediates gene transfer into leukemia cells with high efficiency. Ectopic over-expression of IL-24 has significant growth inhibition and differentiation inducement effects on these cells. Treatment with Ad.RGD-IL-24 could potentially induce leukemia cells' cell-cycle arrest. In addition, IL-24 expression could significantly induce apoptosis of the THP-1 cells. Ad.RGD-IL-24 had a potent effect on the up-regulation of the expression of GRP78/Bip, GADD34 and Bax, down-regulation of the expression of Bcl-2 and Mcl-1, and induced the activation of Caspase-3, which may be responsible for its apoptosis-inducing effect on THP-1 cells. Furthermore, IL-24 expression could retard transplanted leukemia cell tumor growth in vivo in athymic nude mice. These findings showed the marked antitumor activity of IL-24 and provided potential perspectives in designing therapeutic or vaccine strategies in immuno-gene therapy of myeloid leukemia.
Assuntos
Adenoviridae/genética , Interleucinas/genética , Leucemia Mieloide/terapia , Oligopeptídeos/genética , Animais , Apoptose , Ciclo Celular , Diferenciação Celular , Linhagem Celular Tumoral , Chaperona BiP do Retículo Endoplasmático , Terapia Genética , Humanos , Leucemia Mieloide/patologia , Camundongos Nus , RNA Mensageiro/metabolismo , Carga TumoralRESUMO
The inhibitor of growth (ING) family proteins have been defined as candidate tumor suppressors. ING4 as a novel member of ING family has potential tumor-suppressive effects via multiple pathways. However, the therapeutic effect of adenovirus-mediated ING4 (Ad-ING4) gene transfer in human osteosarcoma is still unknown. In this study, we explored the in vitro and in vivo antitumor activity of Ad-ING4 in human osteosarcoma and its potential mechanism using a MG-63 human osteosarcoma cell line. We demonstrated that Ad-ING4 induced significant growth inhibition and apoptosis, upregulated the expression of P21, P27 and Bax, downregulated the Bcl-2 expression and activated Caspase-3 in MG-63 human osteosarcoma cells. Moreover, intratumoral injections of Ad-ING4 in athymic nude mice bearing MG-63 human osteosarcoma tumors significantly suppressed osteosarcoma xenografted tumor growth, increased the expression of P21, P27 and Bax, reduced the Bcl-2 and CD34 expression and microvessel density (MVD) in tumors. This retarded MG-63 osteosarcoma growth in vitro and in vivo in an athymic nude mouse model elicited by Ad-ING4 was closely associated with the increase in the expression of cell cycle-related molecules P21 and P27, decrease in the ratio of anti- to pro-apoptotic molecules Bcl-2/Bax followed by the activation of Caspase-3 leading to apoptosis via intrinsic apoptotic pathways, and the inhibition of tumor angiogenesis. Thus, our results indicate that Ad-ING4 is a potential candidate for human osteosarcoma gene therapy.
Assuntos
Adenoviridae/genética , Apoptose/genética , Neoplasias Ósseas/terapia , Proteínas de Ciclo Celular/genética , Proteínas de Homeodomínio/genética , Neovascularização Patológica/genética , Osteossarcoma/terapia , Proteínas Supressoras de Tumor/genética , Animais , Neoplasias Ósseas/patologia , Caspase 3/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/genética , Inibidor de Quinase Dependente de Ciclina p21/biossíntese , Inibidor de Quinase Dependente de Ciclina p27/biossíntese , Ativação Enzimática/genética , Técnicas de Transferência de Genes , Genes Supressores de Tumor , Terapia Genética/métodos , Humanos , Masculino , Camundongos , Camundongos Nus , Transplante de Neoplasias , Neovascularização Patológica/terapia , Osteossarcoma/patologia , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Transplante Heterólogo , Proteína X Associada a bcl-2/biossínteseRESUMO
The inhibitor of growth (ING) family proteins have been defined as candidate tumor suppressors. ING4 as a novel member of ING family has potential tumor-suppressive effects via multiple pathways. However, the therapeutic effect of adenovirus-mediated ING4 (Ad-ING4) gene transfer in human osteosarcoma is still unknown. In this study, we explored the in vitro and in vivo antitumor activity of Ad-ING4 in human osteosarcoma and its potential mechanism using a MG-63 human osteosarcoma cell line. We demonstrated that Ad-ING4 induced significant growth inhibition and apoptosis, upregulated the expression of P21, P27 and Bax, downregulated the Bcl-2 expression and activated Caspase-3 in MG-63 human osteosarcoma cells. Moreover, intratumoral injections of Ad-ING4 in athymic nude mice bearing MG-63 human osteosarcoma tumors significantly suppressed osteosarcoma xenografted tumor growth, increased the expression of P21, P27 and Bax, reduced the Bcl-2 and CD34 expression and microvessel density (MVD) in tumors. This retarded MG-63 osteosarcoma growth in vitro and in vivo in an athymic nude mouse model elicited by Ad-ING4 was closely associated with the increase in the expression of cell cycle-related molecules P21 and P27, decrease in the ratio of anti- to pro-apoptotic molecules Bcl-2/Bax followed by the activation of Caspase-3 leading to apoptosis via intrinsic apoptotic pathways, and the inhibition of tumor angiogenesis. Thus, our results indicate that Ad-ING4 is a potential candidate for human osteosarcoma gene therapy.
Assuntos
Adenoviridae/genética , Apoptose/genética , Proteínas de Ciclo Celular/genética , Técnicas de Transferência de Genes , Vetores Genéticos/genética , Proteínas de Homeodomínio/genética , Neovascularização Patológica/genética , Osteossarcoma/genética , Osteossarcoma/patologia , Proteínas Supressoras de Tumor/genética , Animais , Biomarcadores , Linhagem Celular Tumoral , Proliferação de Células , Sobrevivência Celular/genética , Modelos Animais de Doenças , Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Terapia Genética , Humanos , Masculino , Camundongos , Neovascularização Patológica/metabolismo , Neovascularização Patológica/terapia , Osteossarcoma/metabolismo , Osteossarcoma/terapia , Carga Tumoral/genética , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
The smallest gene HBx of Hepatitis B virus (HBV) is recognized as an important viral oncogene (V-oncogene) in the hepatocarcinogenesis. Our previous work demonstrated that RMP is a cellular oncogene (C-oncogene) required for the proliferation of hepatocellular carcinoma (HCC) cells. Here we presented the collaboration between V-oncogene HBx and C-oncogene RMP in the development of HCC. The coexpression of HBx and RMP resulted in the cooperative effect of antiapoptosis and proliferation of HCC cells. In vivo, overexpression of RMP accelerated the growth of HBx-induced xenograft tumors in nude mice and vice versa HBx promoted the growth of RMP-driven xenograft tumors. Although HBx didn't regulate the expression of RMP, HBx and RMP interact with each other and collocalized in the cytoplasm of HCC cells. HBx and RMP collaboratively inhibited the expression of apoptotic factors and promoted the expression of antiapoptotic factors. This finding suggests that HBV may induce, or at least partially contributes to the carcinogenesis of HCC, through its V-oncoprotein HBx interacting with the C-oncoprotein RMP.
Assuntos
Carcinoma Hepatocelular/genética , Vírus da Hepatite B/genética , Neoplasias Hepáticas/genética , Proteínas Oncogênicas Virais/metabolismo , Proteínas Repressoras/metabolismo , Transativadores/metabolismo , Animais , Apoptose/genética , Apoptose/fisiologia , Western Blotting , Proliferação de Células/genética , Primers do DNA/genética , Citometria de Fluxo , Células Hep G2 , Humanos , Imuno-Histoquímica , Imunoprecipitação , Camundongos , Microscopia de Fluorescência , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteínas Virais Reguladoras e AcessóriasRESUMO
Di-(2-ethylhexyl) phthalate (DEHP) has been considered as a widespread environmental persistent organic pollutant and its potential concern on human health is raised by previous studies. In particular, children are more likely to be exposed to DEHP through gastrointestinal route and consequently are more susceptible to DEHP hazards. Some reports have uncovered a positive association between DEHP exposure and an increased prevalence of allergic diseases in infants and juveniles. Allergy is a hypersensitive reaction rooted in imbalanced humoral immunity. T follicular helper cell (Tfh), an important CD4(+) Th cell subset, until recently has been identified as a key player in humoral immune response by modifying B cells functions. Tfh cells are therefore perceived as the therapeutic target of immune disorders. In the present study, focusing on the newly confirmed Tfh cells, we examined the effects of DEHP on humoral immunity and investigated the underlying mechanisms. Using ovalbumin (OVA) sensitization weanling mice model under the condition of gastrointestinal exposure to DEHP, we found that DEHP acted as an immunoadjuvant to augment OVA-specific IgE and IgG1 production, amplified germinal center formation in lymphoid nodule, as well as stimulated the expansion of CD4(+)CXCR5(+)ICOS(+)/CD4(+)CXCR5(+)PD-1(+) Tfh cells and CD19(+)CD138(+)GL7(+) plasma cells. Based on the results of immune adoptive transfusion, DEHP-related anaphylactic response was ascribed to Tfh cells hyperfunction directly. We further proved that DEHP gavage together with OVA sensitization adjuvantly promoted the synthesis of cytokines IL-21 and IL-4 and the expression of transcription factors Bcl-6 and c-Maf in Tfh cells. In conclusion, our study demonstrates that DEHP has adjuvant toxic effects on Tfh cells by synthesizing an excess of cytokines IL-21 and IL-4 via over-expression of transcription factors Bcl-6 and c-Maf, leading to an increasing secretion of allergy-related IgE and IgG1.
Assuntos
Dietilexilftalato/toxicidade , Poluentes Ambientais/toxicidade , Centro Germinativo/efeitos dos fármacos , Hipersensibilidade/etiologia , Imunidade Humoral/efeitos dos fármacos , Ativação Linfocitária/efeitos dos fármacos , Ovalbumina , Subpopulações de Linfócitos T/efeitos dos fármacos , Linfócitos T Auxiliares-Indutores/efeitos dos fármacos , Administração Oral , Transferência Adotiva , Animais , Linfócitos B/efeitos dos fármacos , Linfócitos B/imunologia , Proliferação de Células/efeitos dos fármacos , Proteínas de Ligação a DNA/metabolismo , Dietilexilftalato/administração & dosagem , Modelos Animais de Doenças , Poluentes Ambientais/administração & dosagem , Centro Germinativo/imunologia , Centro Germinativo/metabolismo , Hipersensibilidade/sangue , Hipersensibilidade/imunologia , Imunoglobulina E/sangue , Imunoglobulina G/sangue , Interleucina-4/metabolismo , Interleucinas/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Proto-Oncogênicas c-bcl-6 , Proteínas Proto-Oncogênicas c-maf/metabolismo , Medição de Risco , Fatores de Risco , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismo , Subpopulações de Linfócitos T/transplante , Linfócitos T Auxiliares-Indutores/imunologia , Linfócitos T Auxiliares-Indutores/metabolismo , Linfócitos T Auxiliares-Indutores/transplanteRESUMO
PURPOSE: The aim of this study was to develop a rating scale for the weight management of patients with congestive heart failure (CHF). METHODS: The original pool of items was created through in-depth interviews and a literature review. Scale validity was analyzed based on face validity, content validity, and structure validity. The content validity and structure validity were evaluated. The overall internal consistency reliability were assessed by using Cronbach's alpha and retest reliability test. RESULTS: A total of 190 CHF patients were enrolled but 5 refused. The original 19 items were then refined to a scale of 16 items. The final scale included four factors (weight monitoring, knowledge, confidence, and behaviours related to weight management), which accounted for 58.7% of the variance. Content validity ratio on the content validity was 0.88. The Cronbach's alpha was 0.843 and the retest reliability was 0.833. CONCLUSIONS: The Chinese CHF-related weight management scale developed has high reliability and validity. KEY WORDS: Congestive heart failure; Reliability; Scale; Validity; Weight management.
RESUMO
OBJECTIVE: To evaluate the impact of cytomegalovirus (CMV) and Epstein-Barr virus (EBV) co-activation on the prognosis of allogeneic hematopoietic stem cell transplantation (allo-HSCT) patients. METHODS: We retrospectively analyzed 330 consecutive allo-HSCT patients at First Affiliated Hospital of Soochow University from December 2011 to August 2013. CMV and EBV DNA were regularly monitored by quantitative polymerase chain reaction (PCR) from the engraftment of granuloCyte within one year after transplantation. The incidences of viremia and clinical outcomes were analyzed by χ(2) test and Kaplan-Meier analysis. RESULTS: After a median follow-up period of 16 (7-25) months, a total of 113 (34.2%) patients were identified with CMV viremia (CMV+) alone, 82 (24.8%) with EBV viremia (EBV+) alone and 32 (9.7%) with CMV and EBV co-activation (CMV/EBV+). The proportion of patients undergoing HLA mismatched transplantation and ones with acute graft-versus-host disease (aGVHD) in CMV/EBV+ group was significantly higher than CMV+ group or EBV+ group (78.1% (25/32) vs 58.5% (48/82) or 50.4% (57/113), P = 0.047,0.008; 56.3% (18/32) vs 32.9% (27/82) or 34.5% (39/113) , P = 0.022, 0.026) . The incidence of post-transplant lymphoproliferative disorder (PTLD) was similar to EBV+ group (12.5% (4/32) vs 11.0% (9/82) , P = 0.802) and so did the incidence of CMV disease when compared with CMV+ group (9.4% (3/32) vs 7.1% (8/113) , P = 0.665). The 2-year overall survival (OS) of CMV+, EBV+ and CMV/EBV+ groups was 68.7%, 61.5% and 62.4% respectively. And no significant difference existed between CMV/EBV+ and the other two groups (P = 0.598, 0.717). However, the 6-month non-relapse mortality (NRM) of CMV/EBV+ group was significantly higher than that of CMV+ or EBV+ group (18.7% vs 8.9%, P = 0.036; 18.7% vs 8.1%, P = 0.032). CONCLUSIONS: HLA mismatch transplants and aGVHD are frequent in CMV and EBV co-activation group. When compared with EBV+ or CMV+ patients, the CMV/EBV+ patients have similar incidence of PTLD or CMV disease and 2-year OS.However, the 6-month NRM is significantly higher in CMV/EBV+ group. It suggests that CMV and EBV co-activation is a risk factor for early mortality of allo-HSCT patients.
Assuntos
Citomegalovirus , Transplante de Células-Tronco Hematopoéticas , Herpesvirus Humano 4 , Ativação Viral , Infecções por Citomegalovirus , Infecções por Vírus Epstein-Barr , Doença Enxerto-Hospedeiro , Humanos , Incidência , Estimativa de Kaplan-Meier , Recidiva Local de Neoplasia , Prognóstico , Estudos Retrospectivos , Fatores de Risco , Transplante Homólogo , ViremiaRESUMO
Interleukin-24 (IL-24)/melanoma differentiation-associated gene-7 (mda-7) is a unique cytokine-tumor suppressor that displays ubiquitous antitumor properties and tumor-specific killing activity. Oncostatin M (OSM) is the most active IL-6-type cytokine and inhibits the proliferation of various solid tumor cell lines. Multigene-based combination therapy may be an effective practice in cancer gene therapy. The therapeutic potential of a combination of IL-24 and OSM in treating cancers is still elusive. In this study, we aimed to examine the enhanced antitumor activity of adenovirus-mediated IL-24/OSM tumor suppressor gene cotransfer in human melanoma cells. We constructed an IL-24/OSM bicistronic adenovirus and assessed its combined effect on A375 human melanoma cells in vitro and in vivo by detecting and comparing apoptosis in the bicistronic antioncogene group (Ad-IL-24-OSM) and in the IL-24 or OSM single antioncogene group. We also investigated the possible mechanism underlying this effect. The bicistronic adenovirus-mediated coexpression of IL-24 and OSM induced additive growth suppression and apoptosis and an overlapping effect on the upregulation of p21, p53, Bax, and cleaved caspase-3 in vitro and in vivo. Moreover, Ad-IL-24-OSM treatment additively reduced the expression of CDK4 and cyclin D1 in A375 melanoma cells and the expression of CD34 and Cox-2 in A375 xenograft tumors in athymic nude mice. The enhanced antitumor activity elicited by Ad-IL-24-OSM was closely associated with the activation of the apoptotic pathway and the additive inhibition of tumor angiogenesis. Therefore, our results indicate that cancer gene therapy combining two or more tumor suppressors, such as IL-24 and OSM, may constitute a novel and effective therapeutic strategy for treating malignant melanoma and other cancers.
Assuntos
Adenoviridae/genética , Terapia Genética/métodos , Interleucinas/genética , Melanoma/genética , Melanoma/terapia , Oncostatina M/genética , Animais , Apoptose/genética , Linhagem Celular Tumoral , Citometria de Fluxo , Humanos , Imuno-Histoquímica , Interleucinas/biossíntese , Masculino , Melanoma/irrigação sanguínea , Melanoma/patologia , Camundongos , Camundongos Nus , Neovascularização Patológica/genética , Neovascularização Patológica/terapia , Oncostatina M/biossíntese , Plasmídeos/administração & dosagem , Plasmídeos/genética , Distribuição Aleatória , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
RMP has been shown to function in the transcription regulation through association with RNA polymerase (RNAP) II subunit RPB5. It also has been shown to be required for the proliferation of hepatocellular carcinoma (HCC) cells with an antiapoptotic property. In this article, we further demonstrate that RMP displays distinct features in HCC cells compared with normal hepatic cells. RMP expression is remarkably increased in various cancer cell lines including HCC cells when compared with normal cells. Depletion of RMP could inhibit the proliferation of HCC cells, but not the normal hepatic cells. RMP significantly prevented apoptosis of HCC cells in SMMC-7721 and HepG2, but had little effect on apoptosis in the normal hepatic cells. The mechanisms of RMP's distinct features rely on different responsive expressions of apoptosis factors induced by RMP in HCC and hepatic cells. Either overexpression or depletion of RMP significantly affected the expression of apoptosis factors in HCC cells. However, normal hepatic cells showed a tendency to resist RMP for the regulation of apoptosis. In the clinical samples, the increased expression of RMP in HCCs was also observed when compared with the matched non-tumor tissues from 30 HCC patients. The different expression levels of and distinct responses to RMP between HCC and hepatic cells suggest that RMP might serve as not only a biomarker for the diagnosis of HCC, but also a potential target for the HCC therapy.
Assuntos
Carcinoma Hepatocelular/patologia , Proliferação de Células , Peptídeos e Proteínas de Sinalização Intracelular/fisiologia , Neoplasias Hepáticas/patologia , Fígado/citologia , Sequência de Bases , Células Cultivadas , Primers do DNA , Humanos , Reação em Cadeia da Polimerase em Tempo Real , Proteínas Repressoras , Células Tumorais CultivadasRESUMO
Tumor suppressor melanoma differentiation-associated gene-7/interleukin-24 (mda-7/IL-24) has been extensively regarded as an anti-oncogene; however, that whether IL-24, as a member of IL-10 family, is involved in cancer pain was seldom reported before. In this study, we found that IL-24 mediated by adenovirus could significantly increase the plantar mechanical pain threshold in both operation side and contralateral side of the animal models, which were established by injecting 5×103 Walker 256 rat breast cancer ascitic tumor cells into rats' tibia bone medullary canals; IL-24 could also suppress in vitro Walker 256 cells growth by inducing cell apoptosis. Pathologically, IL-24 could protect bone trabecula and substantia corticalis ossium from being completely destructed. Enzyme-linked immunosorbent assay (ELISA) showed that IL-24 treatment could increase the ß-endorphin levels and decrease the IL-6 concentration in plasma of animals. Our study indicated that IL-24 has a potential treatment effect on cancers not only by inhibiting tumor proliferation, but also by the promotion of ß-endorphin synthesis, inhibition of IL-6 secretion to relieve cancer pain.
Assuntos
Hiperalgesia/genética , Interleucinas/genética , Neoplasias/genética , Animais , Apoptose/genética , Linhagem Celular Tumoral , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Expressão Gênica , Hiperalgesia/metabolismo , Interleucinas/metabolismo , Neoplasias/metabolismo , Neoplasias/patologia , Limiar da Dor , RatosRESUMO
This study was aimed to establish a model for detecting the donor chimerism rate following the multi-donor hematopoietic stem cell transplantations, and simplify its calculation method. Patients with hematologic disease receiving allogeneic hematopoietic stem cell transplantation including single-donor and multi-donor were selected in this study and the donor cell chimerism rates were detected, using STR-PCR combined with capillary electrophoresis. The results indicated that the peaks of the sister alleles coming from the same individual were confirmed to have the approximate areas and can be replaced each other in the situation of mixed chimerism. In the calculation model, the value between reference chimerism and approximate chimerism have no significant difference using the hypothetical peak areas, and the result was confirmed to be accepted basing on typical measurement error between sister alleles (5% - 20%). It is concluded that the areas of share peaks can be replaced by non-share peaks and this conclusion can be used to calculate the double-donor CHM (DD-CHM)(%). Compared to the D alleles, R alleles show more strategic importance because it can lead to more accurate result and allowed simplifying the arithmetic calculations for DD-CHM(%).
Assuntos
Transplante de Células-Tronco Hematopoéticas , Quimeras de Transplante/genética , Alelos , Eletroforese Capilar , Humanos , Reação em Cadeia da Polimerase , Período Pós-Operatório , Doadores de Tecidos , Transplante HomólogoRESUMO
In vitro expansion of transplantable hematopoietic stem cells (HSCs) is a very promising approach for different clinical applications. We have recently developed a new culture system that facilitates in vitro expansion of transplantable cord blood HSCs. In our study, we constructed a recombinant adenovirus Ad-GFP/human leukemia inhibitory factor (hLIF) expressing hLIF. The hLIF gene was delivered into human embryo lung fibroblast cell line WI-38 via infection with Ad-GFP/hLIF. Then, the transgenic cells were cultured on regenerated silk fibroin (SF) films as feeder layer cells for expansion of cord blood CD34(+) cells. Our results showed that the hLIF transgenic WI-38 cells cultured on SF could express hematopoiesis-related cytokines at higher levels compared with control groups. The hLIF-expressing feeder layer cells cultured on SF in combination with cytokines more efficiently expanded CD34(+) cells and CD34(+) CD38(-) cells. The percentages of adhesion molecules on the expanded CD34(+) cells in transgenic feeder layer cells cultured on SF were higher than those of control groups. Interestingly, the migration rate assessed by transwell assay was also significantly higher than those of control groups, which suggests that transgenic feeder layer cells cultured on SF has powerful ability to maintain the homing capacity of expanded CD34(+) cells.
Assuntos
Sangue Fetal/citologia , Fibroínas , Fator Inibidor de Leucemia/genética , ADP-Ribosil Ciclase 1/metabolismo , Antígenos CD34/metabolismo , Materiais Biocompatíveis , Linhagem Celular , Proliferação de Células , Técnicas de Cocultura , Transplante de Células-Tronco de Sangue do Cordão Umbilical , Citocinas/biossíntese , Células Alimentadoras/metabolismo , Sangue Fetal/imunologia , Células-Tronco Hematopoéticas/citologia , Células-Tronco Hematopoéticas/imunologia , Humanos , Teste de Materiais , Glicoproteínas de Membrana/metabolismo , Proteínas Recombinantes/genéticaRESUMO
Interleukin-6 (IL6) is a pleiotropic inflammatory cytokine, which is implicated in the development and progression of several types of cancer. The -174G/C polymorphism of the IL6 gene controls serum levels of IL6 and may be associated with cancer risk, but the results from the published studies on the association between this polymorphism and cancer risk are conflicting. A comprehensive meta-analysis was conducted to assess the association of IL6 -174G/C with cancer risk. Studies were identified by searches of MEDLINE and HuGE Published Literature databases, with no restrictions. An eligible 83 articles involving 44,735 cancer patients and 60,747 controls were included. Combined odds ratios (ORs) and 95% confidence intervals (CIs) were used to assess the strength of the association between the IL6 -174 G/C polymorphism and cancer risk. Potential sources of heterogeneity were explored by meta-regression and sensitivity analysis. Overall, the IL6 -174G/C polymorphism was not significantly associated with cancer risk. However, cancer risk was increased for individuals with the CC genotype compared to those carrying the GG genotype in African populations (OR=1.83, 95% CI 1.26-2.67, P=0.002), but not in Caucasian populations (OR=1.00, 95% CI 0.92-1.08, P=0.938). The present meta-analysis provides the first evidence of the ethnic-specific association of the IL6 -174G/C polymorphism with cancer risk. Further investigations with a large number of cases and controls are required to confirm the associations between this polymorphism and cancer in Africans.
RESUMO
OBJECTIVE: To evaluate the prevalence and distribution of C-kit, NPM1 and FLT3 gene mutations in patients with acute myeloid leukemia (AML), and to analyze the relationship between the gene mutations and their prognosis. METHODS: Mutations in exon 8 and 17 of C-kit gene, exon 12 of NPM1 gene, exon 20 of FLT3-TKD gene, and exon 14/15 of FLT3-ITD gene were detected by direct sequencing. Clinical data was collected and followed up if the patient had accepted treatment in our hospital. RESULTS: Among the 656 AML patients, mutations in C-kit exon 8 were found in 6 patients (0.9%), C-kit exon 17 in 33 (5.0%), NPM1 in 169 (25.8%), FLT3-TKD in 46 (7.1%), and FLT3-ITD in 178 (27.1%). Six subtypes of mutations were detected in C-kit exon 8, 8 in C-kit exon 17, 11 in FLT3-TKD, 15 in NPM1, of which 5 were not reported before. C-kit exon 17 mutations were more frequently detected in patients with t(8;21) and exon 8 in patients with inv(16) cytogenetic abnormality. No other gene mutations except FLT3 were detected in M(3) patients. NPM1 and ITD mutations were often detected in individuals with normal cytogenetics or M(5) and M(1) of FAB classification, and accompanied with high white blood cell counts in peripheral blood, high blast counts in bone marrow and low CD34 expression. The older the patients were when diagnosed, the more gene mutations and the higher white blood cell count were detected. More mutations were found in individuals with normal karyotype than that with other karyotypes. It appeared that FLT3-ITD was significantly associated with shorter overall survival (OS) (P = 0.004), NPM1 was not significantly associated with OS, but NPM1(+)/ITD(-) patients had the longest OS. CONCLUSIONS: Our results showed that the mutation types and amounts had particular distribution in MICM subtypes, and were associated with white blood cell counts in peripheral blood, blast counts in bone marrow and prognosis. Especially for patients with normal karyotype, the genetic mutations could be new molecule marker.
Assuntos
Leucemia Mieloide Aguda/diagnóstico , Leucemia Mieloide Aguda/genética , Proteínas Nucleares/genética , Proteínas Proto-Oncogênicas c-kit/genética , Tirosina Quinase 3 Semelhante a fms/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Povo Asiático/genética , Análise Mutacional de DNA , Feminino , Humanos , Cariotipagem , Masculino , Pessoa de Meia-Idade , Mutação , Nucleofosmina , Prognóstico , Adulto JovemRESUMO
Pancreatic cancer has a poor prognosis, even with surgery. ING4 is a member of the inhibitor of growth (ING) tumor suppressor family that has potent inhibitory effects on a variety of tumors; meanwhile, radiotherapy is a common adjunctive therapy for pancreatic cancer. Prior to this study, the effectiveness of a combination of ING4 gene-therapy and radiotherapy against pancreatic cancer had been unknown. In this study, we demonstrated that either ING4 or (125)I radiotherapy treatment could induce Panc-1 pancreatic cancer cell growth suppression and apoptosis in vitro. Furthermore, both treatments inhibited tumor growth and angiogenesis of Panc-1 pancreatic cancer subcutaneously xenografted in vivo. Moreover, the combination therapy had a synergistic effect.
Assuntos
Proteínas de Ciclo Celular/genética , Proteínas de Homeodomínio/genética , Radioisótopos do Iodo/farmacologia , Neoplasias Pancreáticas/terapia , Proteínas Supressoras de Tumor/genética , Adenoviridae/genética , Animais , Antígenos CD34/metabolismo , Apoptose/genética , Apoptose/efeitos da radiação , Proteínas de Ciclo Celular/biossíntese , Linhagem Celular Tumoral , Terapia Combinada , Terapia Genética , Proteínas de Homeodomínio/biossíntese , Humanos , Camundongos , Camundongos Nus , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/metabolismo , Neoplasias Pancreáticas/radioterapia , Antígeno Nuclear de Célula em Proliferação/metabolismo , Distribuição Aleatória , Proteínas Supressoras de Tumor/biossíntese , Fator A de Crescimento do Endotélio Vascular/metabolismo , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
BACKGROUND: Quinacrine (QC), an antimalarial drug, has been shown to possess anticancer effect both in vitro (cancer cell lines) and in vivo (mouse models). In the cancer cells, QC can simultaneously suppress nuclear factor-κB and activate p53 signaling, which results in the induction of the apoptosis in these cells. However, the experimental results come from a few limited cancer cell lines, and the detailed mechanisms remain unknown. OBJECTIVE: This study investigated the tumor-killing effects of QC on gastric cancer cells as well as underlying molecular pathways. METHODS: SGC-7901 cells were treated with or without QC at different concentrations for 24 hours. The effect of QC on the inhibition of SGC-7901 cell proliferation was assessed by Cell Counting Kit-8 assay. Apoptosis was detected by examining nuclear morphology and quantifying phosphatidylserine externalization. Alterations in cellular morphology were analyzed by laser scanning confocal microscopy for fluorescent analysis. Cell cycle analysis was performed by propidium iodide (PI) staining and flow cytometry. The enzyme activity changes of caspase-3 were detected by colorimetry expression method. Western blot analysis was used to detect the changes in the protein level of Bax, Bc1-2, p53, and cytochrome c in cytosol of SGC-7901 cells. RESULTS: Our results showed that QC could significantly inhibit the growth of SGC-7901 cells in a dose-dependent manner, with the IC50 mean (SD) value of 16.18 (0.64) µM, compared with nontreated controls. QC treatment (15 µM) could also induce apoptosis in SGC-7901 cells (26.30% [5.31%], compared with control group of 3.37% [0.81%]; P < 0.01), and the increasing phosphatidylserine level and the accumulation of chromatin nucleation in QC-treated cells provided further evidence. In addition, cell cycle analysis with PI staining showed that a significant S enriches, increasing from 12.00% (1.24%) (control) to 20.94% (2.40%) (QC treatment) (P < 0.01). Furthermore, increased activities of caspase-3 (increasing from 0.108 [0.019] to 0.628 [0.068]; P < 0.01) were observed in SGC-7901 cells treated with 15 µM QC. Western blot analysis showed that QC treatment significantly increased the levels of proapoptotic proteins, including cytochrome c, Bax, and p53, and decreased the levels of antiapoptotic protein Bcl-2, thus shifting the ratio of Bax/Bcl-2 in favor of apoptosis. CONCLUSIONS: Our findings suggest that QC can significantly inhibit cell growth and induce apoptosis in SGC-7901 cells, which involves p53 upregulation and caspase-3 activation pathway.