RESUMEN
17ß-Hydroxysteroid dehydrogenases (17ß-HSDs) are multifunctional enzymes involved in the metabolism of steroids, fatty acids, retinoids and bile acid. In this study, two novel types of 17ß-HSDs (named as MgHsd17b10 and MgHsd17b12) were cloned from Mytilus galloprovincialis by using rapid amplification of cDNA ends (RACE) approaches. Sequence analysis showed that MgHsd17b10 and MgHsd17b12 encoded a polypeptide of 259 and 325 amino acids, respectively. Phylogenetic analysis revealed that MgHsd17b10 and MgHsd17b12 were evolutionarily clustered with other invertebrate 17ß-HSD type 10 and 17ß-HSD type 12 homologues. The MgHsd17b10 and MgHsd17b12 transcripts could be detected in all examined tissues with higher expression levels in digestive glands and gonad. After exposed to endocrine disrupting chemicals (Bisphenol A or 2,2',4,4'-tetrabromodiphenyl ether), the expression of MgHsd17b10 and MgHsd17b12 transcripts was both down-regulated in digestive glands. These findings suggest that MgHsd17b10 and MgHsd17b12 perhaps play an important role in the endocrine regulation of M. galloprovincialis.
Asunto(s)
17-Hidroxiesteroide Deshidrogenasas/genética , Compuestos de Bencidrilo/toxicidad , Disruptores Endocrinos/toxicidad , Éteres Difenilos Halogenados/toxicidad , Mytilus/genética , Fenoles/toxicidad , Contaminantes Químicos del Agua/toxicidad , Animales , Secuencia de Bases , ADN Complementario/genética , Regulación de la Expresión Génica/efectos de los fármacos , Datos de Secuencia Molecular , Filogenia , ARN Mensajero/metabolismo , Análisis de Secuencia de ADNRESUMEN
In this work, behaviors of positively-charged AuNRs in a highly metastatic tumor cell line MDA-MB-231 are examined based on UV-vis-NIR absorption spectroscopy in combination with inductively coupled plasma mass spectrometry (ICP-MS), transmission electron microscopy (TEM) and dark-field microscopic observation. It is found that characteristic surface plasmon resonance (SPR) peaks of AuNRs can be detected using spectroscopic method within living cells that have taken up AuNRs. The peak area of transverse SPR band is shown to be proportionally related to the amount of AuNRs in the cells determined with ICP-MS, which suggests a facile and real time quantification method for AuNRs in living cells. The shape of longitudinal SPR band in UV-vis-NIR spectrum reflects the aggregation state of AuNRs in the cells during the incubation period, which is proved by TEM and microscopic observations. Experimental results reveal that AuNRs are internalized by the cells rapidly; the accumulation, distribution and aggregation of AuNRs in the cells compartments are time and dose dependent. The established spectroscopic analysis method can not only monitor the behaviors of AuNRs in living cells but may also be helpful in choosing the optimum laser stimulation wavelength for anti-tumor thermotherapy.
Asunto(s)
Neoplasias de la Mama/patología , Oro/química , Nanopartículas del Metal/química , Espectrofotometría Ultravioleta/métodos , Espectroscopía Infrarroja Corta/métodos , Calibración , Línea Celular Tumoral , Medios de Cultivo/metabolismo , Femenino , Humanos , Rayos Láser , Lisosomas/química , Espectrometría de Masas/métodos , Microscopía Electrónica de Transmisión/métodos , Modelos Biológicos , Metástasis de la Neoplasia , Neoplasias/metabolismo , Neoplasias/patología , Resonancia por Plasmón de Superficie , Agua/químicaRESUMEN
Mesoporous silica-coated gold nanorods (Au@SiO(2)) are developed as a promising and versatile theranostic platform for cancer treatment. Intracellular localization of Au@SiO(2) is visualized through two-photon imaging. With doxorubicin hydrochloride loaded, Au@SiO(2)-DOX show two light-mediated therapeutic modes: low power density laser-triggered drug release for chemotherapy, and high power density laser-induced hyperthermia, which suggest the potential for in-vivo applications.