RESUMEN
Jade-1 is originally identified by the yeast two-hybrid system as a protein partner of von Hippel-Lindau (pVHL) tumor suppressor, a well-known renal tumor suppressor. In cellular signaling pathways, many upstream Jade-1 regulators, such as pVHL, CK1α, PC1, and NPHP4, can control its activity by stabilization, phosphorylation, and nuclear translocation. Numerous downstream effectors, including ß-catenin, AKT, p21, and Bcl-2, are well modulated by Jade-1, which mainly regulates cell proliferation and apoptosis. Jade-1 is also deemed to be a candidate of transcriptional co-activator associated with histone acetyltransferase (HAT) activity. This review focuses on the anticancer role of Jade-1 in clear cell renal carcinoma and the inhibitory effect of Jade-1 on cystic renal diseases. This review aims to provide a basis of disease prevention or therapy.
Asunto(s)
Carcinoma de Células Renales/metabolismo , Proteínas de Homeodominio/metabolismo , Neoplasias Renales/metabolismo , Proteínas Supresoras de Tumor/metabolismo , Animales , Humanos , Enfermedades Renales Quísticas/metabolismoRESUMEN
AIM: To determinate the RPA1 expression in esophageal carcinoma and the paired tumor-adjacent tissue, and to explore the influence of RPA1 on radiosensitivity of esophageal carcinoma TE-1 cells. METHODS: Firstly, the RPA1 expression of 40 cases esophageal carcinoma and their adjacent tissues were detected by immunohistochemistry. Secondly, The esophageal carcinoma cell subline-radiation resistance model (TE-1R) was constructed by radiation-induction, the RPA1 expression and proliferation activity of TE-1 and TE-1R cells were detected by Western blot and MTT assay respectively. After radiation, the expression of RPA1 and cell apoptosis were detected by Western blot and FACS respectively. Cell clone formation and survival rate were detected by clonogenic assay. Thirdly, Inhibiting RPA1 expression by siRNA in TE-1 cells, the expression of RPA1 was detected by RT-PCR and Western blot, Cell proliferation inhibition ratio and cell apoptosis after radiation were detected by MTT assay and FACS respectively. RESULTS: The RPA1 expression in esophageal carcinoma was significantly higher than that in the tumor-adjacent tissues, which was associated with tumor invasion and lymph node metastasis. The RPA1 expression in TE-1R cells was higher than that in TE-1 cells, while the proliferation activity of TE-1R cells was lower than that of TE-1 cells, and the apoptosis rate of TE-1R cells after radiation was less than that of TE-1 cells. In addtion, the clone formation and survival rate of TE-1R cells were higher than that of TE-1 cells. Moreover, inhibiting RPA1 expression by siRNA-RPA1 could promoted proliferation inhibition ratio and apoptosis rate of TE-1 cells after radiation. CONCLUSION: The over-expression of RPA1 in esophageal carcinoma was related with progression and metastasis. Moreover, radiation induced the excessive expression RPA1 in TE-1 cells, and the radiosensitivity of TE-1R cells was less than that of TE-1 cells. Furthermore, inhibiting RPA1 expression could increase radiosensitivity of TE-1 cells. Overall, RPA1 could influence radiosensitivity and might be one important mechanism of radiation resistance in TE-1 cells.
Asunto(s)
Neoplasias Esofágicas/metabolismo , Tolerancia a Radiación , Proteína de Replicación A/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Apoptosis , Línea Celular Tumoral , Proliferación Celular , Supervivencia Celular , Neoplasias Esofágicas/genética , Neoplasias Esofágicas/patología , Neoplasias Esofágicas/radioterapia , Femenino , Humanos , Masculino , Persona de Mediana Edad , ARN Interferente Pequeño/genéticaRESUMEN
It has been shown that interleukin 18 (IL-18) exerts antitumor activity. In this study, we investigated whether oncolytic adenovirus-mediated gene transfer of IL-18 could induce strong antitumor activity. A tumor-selective replicating adenovirus expressing IL-18 (ZD55-IL-18) was constructed by insertion of an IL-18 expression cassette into the ZD55 vector, which is based on deletion of the adenoviral E1B 55-kDa gene. It has been shown that ZD55-IL-18 exerted a strong cytopathic effect and significant apoptosis in tumor cells. ZD55-IL-18 significantly decreased vascular endothelial growth factor and CD34 expression in the melanoma cells. Treatment of established tumors with ZD55-IL-18 showed much stronger antitumor activity than that induced by ZD55-EGFP (enhanced green fluorescent protein) or Ad-IL-18. These data indicated that oncolytic adenovirus expressing IL-18 could exert potential antitumor activity through inhibition of angiogenesis and offer a novel approach to melanoma therapy.
Asunto(s)
Adenoviridae/fisiología , Interleucina-18/genética , Melanoma/terapia , Viroterapia Oncolítica/métodos , Adenoviridae/genética , Adenoviridae/metabolismo , Animales , Apoptosis/efectos de los fármacos , Apoptosis/genética , Línea Celular Tumoral , Terapia Genética/métodos , Interleucina-18/biosíntesis , Masculino , Melanoma/virología , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Neovascularización Patológica/terapia , Neovascularización Patológica/virología , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
It has been demonstrated that growth hormone (GH) transgenic fish often posses a trait for fast growth. Here, we investigated the growth of F(4)'all-fish' GH transgenic carp Cyprinus carpio and their serum GH levels for a year. The results showed that F(4) all-fish GH transgenic carp were significantly larger in body mass (c. two-fold, P < 0.001) and body length (c. 1.3 fold, P < 0.001), compared with the non-transgenic group. The discrepancy of serum GH levels between the transgenic carp group and control group is 54 fold, when the water temperature was 12-34 degrees C. When the water temperature decreased to 3.5 degrees C in January, the discrepancy was 256 fold. The serum GH level of the transgenic group was relatively constant, while that of control varied greatly based on month and water temperature. The changes of growth rates between the transgenic group and the control group were similar for a year. Taken together, the results indicated that F(4) all-fish GH transgenic carp had not only higher and constant serum GH levels but also a significant fast-growing effect, compared with the control. To our knowledge, this is the first report on a one-year investigation of growth trait and serum growth hormone level in F(4) all-fish GH transgenic carp.
Asunto(s)
Tamaño Corporal/genética , Carpas/sangre , Carpas/crecimiento & desarrollo , Hormona del Crecimiento/sangre , Animales , Animales Modificados Genéticamente , Femenino , Hormona del Crecimiento/genética , Masculino , TemperaturaRESUMEN
RNA interference (RNAi) has been proved to be a powerful tool for gene knockdown purpose and holds great promise for the treatment of cancer. Our previous study demonstrated that the reduction of Ki-67 expression by means of chemically synthesized siRNAs and shRNAs expressed from plasmid resulted in proliferation inhibition in human renal carcinoma cells. In this study, we constructed a novel oncolytic adenovirus-based shRNA expression system, ZD55-Ki67, and explored ZD55-Ki67-mediated RNAi for Ki-67 gene silencing. Our results showed that ZD55-Ki67 could induce silencing of the Ki-67 gene effectively, allow for efficient tumor-specific viral replication and induce the apoptosis of tumor cells effectively in vitro and in nude mice. We conclude that combining shRNA gene therapy and oncolytic virotherapy can enhance antitumor efficacy as a result of synergism between CRAd oncolysis and shRNA antitumor responses.
Asunto(s)
Adenoviridae , Apoptosis , Terapia Genética , Antígeno Ki-67/biosíntesis , Neoplasias Renales/terapia , Proteínas de Neoplasias/sangre , Viroterapia Oncolítica , Virus Oncolíticos , ARN Mensajero/antagonistas & inhibidores , ARN Neoplásico/antagonistas & inhibidores , ARN Interferente Pequeño , Animales , Apoptosis/genética , Línea Celular Tumoral , Técnicas de Silenciamiento del Gen/métodos , Humanos , Antígeno Ki-67/genética , Neoplasias Renales/genética , Neoplasias Renales/metabolismo , Ratones , Ratones Desnudos , Proteínas de Neoplasias/antagonistas & inhibidores , Proteínas de Neoplasias/genética , Interferencia de ARN , ARN Mensajero/genética , ARN Mensajero/metabolismo , ARN Neoplásico/genética , ARN Neoplásico/metabolismo , ARN Interferente Pequeño/genética , Replicación Viral/genéticaRESUMEN
The crosstalk between naive nucleus and maternal factors deposited in egg cytoplasm before zygotic genome activation is crucial for early development. In this study, we utilized two laboratory fishes, zebrafish (Danio rerio) and Chinese rare minnow (Gobiocypris rarus), to obtain mutual crossbred embryos and examine the interaction between nucleus and egg cytoplasm from different species. Although these two types of crossbred embryos originated from common nuclei, various developmental capacities were gained due to different origins of the egg cytoplasm. Using cDNA amplified fragment length polymorphism (cDNA-AFLP), we compared transcript profiles between the mutual crossbred embryos at two developmental stages (50%- and 90%-epiboly). Three thousand cDNA fragments were generated in four cDNA pools with 64 primer combinations. All differentially displayed transcript-derived fragments (TDFs) were screened by dot blot hybridization, and the selected sequences were further analyzed by semi-quantitative RT-PCR and quantitative real-time RT-PCR. Compared with ZR embryos, 12 genes were up-regulated and 12 were down-regulated in RZ embryos. The gene fragments were sequenced and subjected to BLASTN analysis. The sequences encoded various proteins which functioned at various levels of proliferation, growth, and development. One gene (ZR6), dramatically down-regulated in RZ embryos, was chosen for loss-of-function study; the knockdown of ZR6 gave rise to the phenotype resembling that of RZ embryos.
Asunto(s)
Análisis del Polimorfismo de Longitud de Fragmentos Amplificados/veterinaria , Cyprinidae/embriología , Proteínas de Peces/metabolismo , Perfilación de la Expresión Génica , Pez Cebra/embriología , Animales , Cruzamientos Genéticos , Cyprinidae/genética , ADN Complementario/genética , Proteínas de Peces/genética , Regulación del Desarrollo de la Expresión Génica/fisiología , Silenciador del Gen , Transcripción Genética , Pez Cebra/genéticaRESUMEN
Cross-species nuclear transfer (NT) has been used to retain the genetic viability of a species near extinction. However, unlike intra-species NT, most embryos produced by cross-species NT were unable to develop to later stages due to incompatible nucleo-cytoplasmic interactions between the donor nuclei and the recipient cytoplasm from different species. To study the early nucleo-cytoplasmic interaction in cross-species NT, two laboratory fish species (zebrafish and rare minnow) from different subfamilies were used to generate cross-subfamily NT embryos in the present study. Suppression subtractive hybridization (SSH) was performed to screen out differentially expressed genes from the forward and reverse subtractive cDNA libraries. After dot blot and real-time PCR analysis, 80 of 500 randomly selective sequences were proven to be differentially expressed in the cloned embryos. Among them, 45 sequences shared high homology with 28 zebrafish known genes, and 35 sequences were corresponding to 22 novel expressed sequence tags (ESTs). Based on functional clustering and literature mining analysis, up- and down-regulated genes in the cross-subfamily cloned embryos were mostly relevant to transcription and translation initiation, cell cycle regulation, protein binding, etc. To our knowledge, this is the first report on the determination of genes involved in the early development of cross-species NT embryos of fish.
Asunto(s)
Clonación de Organismos , Cyprinidae/fisiología , Embrión no Mamífero/fisiología , Regulación del Desarrollo de la Expresión Génica/fisiología , Pez Cebra/fisiología , Animales , Secuencia de Bases , Análisis por Conglomerados , Cyprinidae/embriología , Cyprinidae/genética , Cartilla de ADN/química , Etiquetas de Secuencia Expresada , Regulación del Desarrollo de la Expresión Génica/efectos de la radiación , Datos de Secuencia Molecular , Técnicas de Transferencia Nuclear/veterinaria , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinaria , Pez Cebra/embriología , Pez Cebra/genéticaRESUMEN
Our previous studies and the others have strongly suggested that c-Jun N-terminal kinase (JNK) signaling pathway plays a critical role in ischemic brain injury. Here we reported that Tat-JNK binding domain (JBD) of JNK-interacting protein-1 (JIP-1), a smaller 11-mer peptide corresponding to residues 153-163 of murine JIP-1 conjugated to Tat peptide, perturbed the assembly of JIP-1-JNK3 complexes, thus inhibiting the activation of JNK3 induced by ischemia/reperfusion in the vulnerable hippocampal CA1 subregion. As a result, Tat-JBD diminished the increased phosphorylation of c-Jun (a nuclear substrate of JNK) and the increased expression of Fas ligand induced by ischemia/reperfusion in the vulnerable hippocampal CA1 subregion. At the same time, through inhibiting phosphorylation of Bcl-2 (a cytosolic target of JNK) and the release of Bax from Bcl-2/Bax dimers, Tat-JBD attenuated Bax translocation to mitochondria and the release of cytochrome c induced by ischemia/reperfusion. Furthermore, the activation of caspase3 and hydrolyzation of poly-ADP-ribose-polymerase induced by brain ischemia/reperfusion were also significantly suppressed by preinfusion of the peptide Tat-JBD. Importantly, Tat-JBD showed neuroprotective effects on ischemic brain damage in vivo, and administration of the peptide after ischemia also achieved the same effects as preinfusion of the peptide did. Thus, our findings imply that Tat-JBD induced neuroprotection against ischemia/reperfusion in rat hippocampal CA1 region via inhibiting nuclear and non-nuclear pathways of JNK signaling. Taken together, these results indicate that Tat-JBD peptide provides a promising therapeutic approach for ischemic brain injury.
Asunto(s)
Lesiones Encefálicas/prevención & control , Proteínas Quinasas JNK Activadas por Mitógenos/antagonistas & inhibidores , Fármacos Neuroprotectores/uso terapéutico , Péptidos/uso terapéutico , Transducción de Señal/efectos de los fármacos , Análisis de Varianza , Animales , Lesiones Encefálicas/etiología , Isquemia Encefálica/complicaciones , Caspasa 3 , Caspasas/metabolismo , Células Cultivadas , Modelos Animales de Enfermedad , Embrión de Mamíferos , Proteína Ligando Fas , Expresión Génica/efectos de los fármacos , Productos del Gen tat/genética , Productos del Gen tat/metabolismo , Hipocampo/citología , Proteínas Quinasas JNK Activadas por Mitógenos/química , Masculino , Glicoproteínas de Membrana/metabolismo , Neuronas/efectos de los fármacos , Péptidos/química , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Ratas , Transducción de Señal/fisiología , Fracciones Subcelulares/efectos de los fármacos , Factores de Necrosis Tumoral/metabolismo , Proteína X Asociada a bcl-2/metabolismo , Receptor fas/metabolismoRESUMEN
Total RNA was extracted from murine hepatocytes, and the cDNA of interleukin 18(IL-18) was amplified by RT-PCR. The cDNA was introduced into the expression vector pJW2 and sequenced. Under heat induction, the recombinant murine IL-18(rmIL-18) was expressed in inclusion bodies in E. coli with the yield accounting for 18% of total bacteria proteins. The inclusion bodies were dissolved with 5 mol/L urea, and rmIL-18 was purified using Sephadex G-100 column chromatography. In the presence of 0.5 mg/L Con A, the purified rmIL-18 showed dose-dependent IFN-gamma-inducing activity in murine splenocytes. The purified rmIL-18 exhibited significant antitumor effects in Kunming mice challenged intraperitoneally (i.p.) with H22 hepatocarcinoma when administered 10 micrograms rmIL-18 i.p. on days 1, 4 after challenge, and the mice survived resisted the rechallenged with H22 cells.