RESUMEN
Grass carp reovirus (GCRV) is the most virulent agent to Grass carp, Ctenopharyngodon idella, and causes a severe infectious disease called hemorrhagic disease of grass carp. Generally, barbel chub, Squaliobarbus curriculus, a genetically closely related species to grass carp, exhibits significant resistance against GCRV infection compared to grass carp. To investigate whether the Toll-like receptor 22 (tlr22) has got a vital role against the GCRV infection, the full cDNA sequence of tlr22 from barbel chub (Sctlr22) was cloned by RACE-PCR, and the structure and expression feature were studied. The complete cDNA sequence of Sctlr22 has a size of 3504 bp, encoding for 960 amino acid residues. Sctlr22 possesses typical structural features of the tlrs family, including 19 leucine rich repeats (LRRs), a transmembrane (TM) and a Toll/interleukin-1 receptor (TIR) domain. Phylogenetic analysis revealed that barbel chub Tlr22 was clustered together with the Tlr22 of grass carp (Citlr22). Structurally, barbel chub Tlr22 have two different structure in LRRs domain and TIR domain with grass carp (Susceptible to GCRV), but was similar to that of Danio rerio and Cyprinus carpio (Resistance to GCRV). Quantitative RT-PCR analysis has shown that Sctlr22 is prominently expressed in immune relevant tissues such as head kidney and spleen. After GCRV infection, Sctlr22 expression level was up-regulated in four tested tissues and the highest expression of Sctlr22 appeared fast and higher than Citlr22. The interferon-ß (ifn-ß) expression level in CIK cells over-expressing fused cDNA encoding the LRR domain of Sctlr22 to the transmembrane and TIR domain of Citlr22 was significantly higher than that cells overexpressing Citlr22 after GCRV infection. The virus titer was significantly reduced compared to Citlr22 over-expressing cells. These results suggested that Sctlr22 seems to play a vital role in the immune response against GCRV.
Asunto(s)
Cyprinidae , Enfermedades de los Peces/genética , Proteínas de Peces/genética , Regulación de la Expresión Génica/inmunología , Inmunidad Innata , Infecciones por Reoviridae/veterinaria , Receptores Toll-Like/genética , Animales , Carpas , Cyprinidae/clasificación , Enfermedades de los Peces/inmunología , Enfermedades de los Peces/virología , Proteínas de Peces/química , Proteínas de Peces/metabolismo , Filogenia , Estructura Terciaria de Proteína , Distribución Aleatoria , Reoviridae/fisiología , Infecciones por Reoviridae/genética , Infecciones por Reoviridae/inmunología , Infecciones por Reoviridae/virología , Receptores Toll-Like/química , Receptores Toll-Like/metabolismoRESUMEN
OBJECTIVE: We investigated the effect of docosahexaenoic acid (DHA) on the invasion and metastasis of ovarian cancer cells (A2780, HO8910, and SKOV-3). METHODS: Cytotoxicity assay was performed to determine the optimal doses of DHA in this experiment. The effects of DHA on invasion ability were assessed by invasion assay. The expressions of messenger RNA and/or proteins associated with invasion or metastasis were detected by quantitative Real Time-Polymerase Chain Reaction or Western blot. The effect of DHA on cell metastasis was assessed in xenograft model of zebrafish. RESULTS: Docosahexaenoic acid and α-linolenic acid could reduce the cell vitalities in dose-dependent manner. However, DHA inhibited the invasion and metastasis of ovarian cancer cells, but α-linolenic acid did not (**P < 0.01). Docosahexaenoic acid could downregulate the expressions of WAVE3, vascular endothelial cell growth factor, and MMP-9, and upregulate KISS-1, TIMP-1, and PPAR-γ, which negatively correlated with cell invasion and metastasis (*P < 0.05). Docosahexaenoic acid restrained the development of subintestinal vessels and cancer cell metastasis in xenograft model of zebrafish (**P < 0.01). CONCLUSIONS: Docosahexaenoic acid inhibited the invasion and metastasis of ovarian cancer cells in vitro and in vivo through the modulation of NF-κB signaling pathway, suggesting that DHA is a promising candidate for ovarian cancer therapy.
Asunto(s)
Ácidos Docosahexaenoicos/farmacología , Neoplasias Ováricas/tratamiento farmacológico , Animales , Animales Modificados Genéticamente , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Regulación hacia Abajo/efectos de los fármacos , Femenino , Humanos , Masculino , Invasividad Neoplásica , Metástasis de la Neoplasia , Neoplasias Ováricas/genética , Neoplasias Ováricas/metabolismo , Neoplasias Ováricas/patología , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Ensayos Antitumor por Modelo de Xenoinjerto , Pez CebraRESUMEN
Glioblastoma, the most common tumor in the brain, has witnessed very little clinical progress over the last decades. Exploring and discovering new therapeutic strategies for glioblastoma has become a critical problem. Harmine (HM), belonging to the beta-carboline alkaloid, is a natural product and isolated from the seeds of Peganum harmala L., which own notable antitumor activity in vitro. However, the poor water solubility and less selectivity of HM severely limit its clinical use. For enhancing its selective ability to tumor cells, we fabricated a kind of protein nanoparticles (BSA-HM NPs), composed of the modified bovine serum albumin (BSA) and HM. It was substantiated through in vitro and in vivo experiment that BSA-HM NPs could predominantly accumulate in tumor tissues and exhibited remarkably enhanced antitumor efficacy. This study provides a promising strategy to improve the bioavailability and avoid side effects of HM as antitumor agents by choosing BSA as carriers.
Asunto(s)
Antineoplásicos , Glioblastoma , Nanopartículas , Antineoplásicos/farmacología , Glioblastoma/tratamiento farmacológico , Harmina/farmacología , Humanos , Albúmina Sérica BovinaRESUMEN
The complete mitochondrial genome sequence of the hybrid of Ctenopharyngodon idella (â) × Squaliobarbus curriculus (â) was determined using PCR-based method. The total length of the mitogenome is 16,609 bp. It contains the typical structure as that of most other vertebrates, including 2 ribosomal RNA genes, 13 protein-coding genes, 22 transfer RNA genes, and 1 non-coding control region (D-loop region). The overall composition of the mitogenome was estimated to be 31.86% for A, 26.08% for T, 26.38% for C, and 15.68% for G, respectively, indicating that an A+T (57.94%)-rich feature occurs in the hybrid mitogenome. Both the termination-associated sequence and three conserved sequence blocks (CSB1, CSB2, and CSB3) were also detected in the D-loop region.
Asunto(s)
Carpas/genética , Cyprinidae/genética , ADN Mitocondrial/genética , Genes Mitocondriales/genética , Genoma Mitocondrial/genética , Hibridación Genética/genética , Animales , Composición de Base , Secuencia de Bases , Secuencia Conservada/genética , Orden Génico/genética , Tamaño del Genoma/genética , Datos de Secuencia Molecular , Análisis de Secuencia de ADNRESUMEN
In this work, we reported the complete mitochondrial DNA sequence of the hybrid of Squaliobarbus curriculus (â) × Ctenopharyngodon idella (â), which was obtained by artificial hybridization. The total length of the mitochondrial genome is 16,616 bp, with the base composition of 31.15% A, 25.02% T, 27.66% C, and 16.17% G. It contains 2 ribosomal RNA genes, 13 protein-coding genes, 22 transfer RNA genes, and a major non-coding control region (D-loop region). The arrangement of these genes is the same as that found in the teleosts. All the protein initiation codons are ATG, except for COX1 that begins with GTG. The complete mitogenome of the hybrid of S. curriculus (â) × C. idella (â) provides an important data set for the study in genetic mechanism.
Asunto(s)
Cyprinidae/genética , ADN Mitocondrial/genética , Genes Mitocondriales/genética , Genoma Mitocondrial/genética , Hibridación Genética/genética , Animales , Composición de Base , Secuencia de Bases , China , Tamaño del Genoma/genética , Datos de Secuencia Molecular , Análisis de Secuencia de ADNRESUMEN
The barbel chub (Squaliobarbus curriculus), a kind of small commercial fish, is widespread in China. In this study, we sequenced the mitochondrial genome of the barbel chub from the Xiangjiang River. The total length of the mitochondrial genome is 16,619 bp, with the base composition of 31.19% A, 25.01% T, 27.68% C, and 16.12% G. The organization and arrangement of these genes are the same as that found in the teleosts, including 2 ribosomal RNA genes, 13 protein-coding genes, 22 transfer RNA genes, and a major noncoding control region (D-loop region). Compared with the S. curriculus collected from Jiangsu province, there were 29 mutation sites in the mitogenome sequence of Xiangjiang S. curriculus. All the mutation sites were transitions and mainly occurred in protein-coding genes (21/29), two mutation sites occurred in transfer RNA, two occurred in ribosomal RNA, and four occurred in D-loop region. Among the 21 mutation sites in protein-coding genes, 6 mutation sites resulted in amino acid mutation in ND2, ATPase6, COX3, ND4, and Cytb genes, while the others were synonymous substitutions. These results indicated that there was genetic variation in different geographical populations of S. curriculus.