RESUMEN
In order to identify genomic changes associated with an etoposide resistance acquisition, we used comparative genomic hybridization (CGH) to compare a human lung adenocarcinoma cell line, A549 wild type, and three sublines, A549-VP1-3, exposed to increasing concentrations of the topoisomerase II inhibitor, VP16. R-banding karyotype, fluorescence in situ hybridization (FISH), and Southern blot for the MLL gene were also performed. The CGH analysis showed that the A549-VP3 cell line shared chemoresistance-specific abnormalities (amplification of 11q23-qter, loss of chromosome 17, and deletions of 2p14-pter and 2q23-q24). FISH analysis confirmed the loss of one chromosome 17 in the three resistant sublines and revealed an increased fragmentation of chromosome 2 in more than two segments, depending on the etoposide concentration. FISH with an MLL gene probe showed additional signals of MLL (from three in the A549-WT to seven in the A549-VP3 cell line) translocated onto several other chromosomes. Southern blot indicated an amplification of the MLL gene, dependent on the etoposide concentration, without gene rearrangement. The CGH results are suggestive of loci that could be associated with the acquisition of an etoposide-chemoresistant phenotype. Deletion of the 2p region has already been reported, without any candidate gene being identified. The role of MLL in leukemogenesis has previously been demonstrated, but its role in the development of other tumors or its significance in the chemoresistance process remains to be elucidated.
Asunto(s)
Adenocarcinoma/genética , Resistencia a Múltiples Medicamentos/genética , Resistencia a Antineoplásicos/genética , Inhibidores Enzimáticos/farmacología , Etopósido/farmacología , Marcadores Genéticos , Neoplasias Pulmonares/genética , Inhibidores de Topoisomerasa II , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/fisiología , Adenocarcinoma/tratamiento farmacológico , Adenocarcinoma/enzimología , Southern Blotting , Humanos , Hibridación Fluorescente in Situ , Cariotipificación , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/enzimología , Hibridación de Ácido Nucleico , Células Tumorales CultivadasRESUMEN
Identification of supernumerary de novo marker chromosomes was considered up to now as difficult and sometimes impossible with classical cytogenetical banding methods. The determination of their chromosomal origin is now easier with fluorescent in situ hybridisation techniques and enables an exact correlation between chromosomal aberration and phenotypic features to be established. The authors describe the use of chromosome painting with chromosome 13 and 18 Whole library DNA probe for identification of supernumerary markers in tow patients with congenital disorders. Cytogenetic examination in the first cave revealed a mosaicism with a ring chromosome 13 but clinical findings were different from the classical "ring 13 syndrome', and chromosome painting revealed in an extra--dicentric 13 chromosome (mos : 47, XX, -13, +r (13) +dic (13) / 46, XX, r (13) / 45, XX, -13 / 48, XX, -13, +r (13), (12) dic (13) / 47, XX, -13, + (2) r (13), R-banding pattern on prometaphases and chromosome painting in the second case confirmed the marker to be a 18 p isochromosome (47, XX, +i (18p)). The feasibility and the usefulness of chromosome painting in ascertainment of the possible genetic significance of markers is discussed.
Asunto(s)
Bandeo Cromosómico/métodos , Cromosomas Humanos Par 13 , Cromosomas Humanos Par 18 , Discapacidad Intelectual/genética , Monosomía , Cromosomas en Anillo , Sondas de ADN , Femenino , Marcadores Genéticos , Humanos , Hibridación Fluorescente in Situ , Lactante , CariotipificaciónRESUMEN
A 18 months old female child with a de novo 16q deletion is described. The clinical findings in this patient are similar to the phenotype first described by Fryns et al. (11) in 16q deletion. The present deletion involves the region 16q11.2-q12.2 suggesting a second critical smallest region of overlap (S.R.O.) more proximal to the centromere than the one previously located in 16q21.
Asunto(s)
Anomalías Múltiples/genética , Aberraciones Cromosómicas/genética , Deleción Cromosómica , Cromosomas Humanos Par 16 , Huesos Faciales/anomalías , Reflujo Gastroesofágico/genética , Cráneo/anomalías , Anomalías Múltiples/diagnóstico , Trastornos de los Cromosomas , Femenino , Reflujo Gastroesofágico/diagnóstico , Hernia Hiatal/diagnóstico , Hernia Hiatal/genética , Humanos , Lactante , Discapacidad Intelectual/diagnóstico , Discapacidad Intelectual/genética , Cariotipificación , Pulgar/anomalíasRESUMEN
Enzyme-linked immuno-filtration assay is carried out on a micropore membrane. This doubly analytical technique permits simultaneous study of antibody specificity by immunoprecipitation and characterisation of antibody isotypes by immuno-filtration with enzyme-labelled antibodies. Recognition of the same T. gondii antigenic constituent by IgG, IgA, IgM or IgE antibodies produces couplets (IgG-IgM; IgG-IgA) or triplets (IgG-IgM-IgA; IgG-IgM-IgE) which identify the functional fractions of the toxoplasmosis antigen. In acquired toxoplasmosis, the persistence of IgM antibody long after infestation puts in question the implication of recent infestation normally linked to detection of this isotype. For sera of comparable titres, comparison of immunological profiles by the method described demonstrates disparities in the composition of the specific antibody content as expressed in international units. Use of the same method to detect IgM antibodies or distinguish between transmitted maternal IgG and IgG antibodies synthesised by the foetus or neonate makes a diagnosis of congenital toxoplasmosis possible in 85% of cases during the first few days of life. With the method described the diagnosis may be made on average 5 months earlier than with classical techniques. In the course of surveillance for latent congenital toxoplasmosis, the appearance of IgM or IgE antibodies raises the possibility of complications (hydrocephalus, chorioretinitis). After cessation of treatment, a rise in IgG antibodies indicating persistence of infection is detected earlier by the present than by classical methods.