RESUMEN
<p><b>OBJECTIVE</b>To investigate the change of expression level of metastasis suppressor gene Kiss-1 in the colorectal cancer cell line SW480 after radiation, and to determine its association with the proliferation and apoptosis of SW480 cells.</p><p><b>METHODS</b>SW480 cells were divided into control group (0 Gy) and study groups (2, 4, 6, 8 Gy). Cells in the study groups were irradiated by 6-MV X-ray radiation for 48 hours. Immunohistochemistry and real-time PCR methods were used to investigate the influence of radiation on Kiss-1 gene expression of SW480. Colony formation assay was used to detect the proliferation of SW480. Flow cytometry-Annexin- V/PI assay was used to observe the change of the apoptosis rate.</p><p><b>RESULTS</b>Compared with the control group, Kiss-1 protein expression increased after radiation of 6, 8 Gy (P<0.05), but no significant changes were observed after radiation of 2, 4 Gy(P>0.05). Kiss-1 gene mRNA level increased after radiation of 2, 4, 6 Gy, while no obvious change was observed for 8 Gy radiation. The apoptosis rates increased for 4, 6, 8 Gy radiation(P<0.05), however, there was no significant difference for 2 Gy radiation (P<0.05).</p><p><b>CONCLUSION</b>Radiation may increase Kiss-1 gene expression in SW480 cells, which results in decreases proliferation and increases apoptosis in residual surviving cells.</p>
Asunto(s)
Humanos , Apoptosis , Línea Celular Tumoral , Proliferación Celular , Neoplasias Colorrectales , Metabolismo , Patología , Kisspeptinas , Genética , Metabolismo , Efectos de la Radiación , ARN Mensajero , Genética , Rayos XRESUMEN
Molecular cloning and nucleotide sequencing of cDNA encoding an orange-spotted grouper (Epinephelus coioides) homolog of Mx ("OsgMx") was conducted and its possible role in fish immunity was analysed. Similar to mammalian Mx, the OsgMx are members of a family of interferon-inducible genes that are expressed by cells in response to nodavirus and iridovirus naturally-infected. Expression of OsgMx mRNA was noticeably upregulated in all tissues by nodavirus naturally-infected grouper. The transcription of OsgMx gene increased 6 h after intramuscular injection of nodavirus experimentally-infected fish and peaked at 72 h in their brains. Analysis of the 5'-flanking sequence of the gene shows that as in pufferfish and zebrafish, the OsgMx promoter contains two potential interferon-stimulated response element (ISRE) responsible for the induction of interferon-inducer polyinosinic-polycytidylic acid (Poly[I:C]). Transient transfection of grouper cells in gfp-reporter gene assays shows that the activation of the grouper Mx promoter fragment by Poly[I:C] is sufficient to allow the expression of green fluorescent protein (GFP). These results may provide a possible regulated pathway against nodavirus.
Asunto(s)
Enfermedades de los Peces/metabolismo , Enfermedades de los Peces/virología , Proteínas de Unión al GTP/genética , Regulación de la Expresión Génica , Nodaviridae , Perciformes/genética , Infecciones por Virus ARN/veterinaria , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , Análisis por Conglomerados , Cartilla de ADN , Proteínas Fluorescentes Verdes , Datos de Secuencia Molecular , Proteínas de Resistencia a Mixovirus , Filogenia , Poli I-C/metabolismo , Infecciones por Virus ARN/metabolismo , Elementos de Respuesta/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinaria , Análisis de Secuencia de ADN/veterinaria , Transfección/veterinariaRESUMEN
Fungi belonging to the Cordyceps species have long been used as food and herbal medicines in Asia and are especially popular as commercially available powdered supplements. Despite this acceptance and use, little is known of the phylogenetic relationships of the genus. Presently, the neighbor-joining method based on the ITS1, 5.8S rRNA, and ITS2 regions was used to construct a phylogenetic tree of 17 Cordyceps isolates. Five major groups were evident. Cordyceps sinensis was less closely related to 15 Cordyceps species but shared a closer relationship with Cordyceps agriota. PCR-single-stranded conformational polymorphism was applied to differentiate seven Cordyceps isolates: five were different from those used to construct the phylogenetic tree, based on differences in the internal spacer 2 (ITS2). The length of ITS2, amplified by primers 5.8SR and ITS4, vary between 334 and 400 bp. This segment could be used for intraspecies classification or detection of mutations and represents potential novel means of identification of this fungal genus in herbal medicines and in quality control applications in the fermentation industry.