RESUMEN

Angioimmunoblastic T-cell Lymphoma (AITL) is one of the most frequent T-cell lymphoma entities. Follicular helper T lymphocytes (TFH) are recognized as the normal cellular counterpart of the neoplastic component. Despite a clonal T-cell feature and few described recurrent cytogenetic abnormalities, a driving oncogenic event has not been identified so far. It has been recently reported that in mice, heterozygous inactivation of Roquin/Rc3h1, a RING type E3 ubiquitine ligase, recapitulates many of the clinical, histological, and cellular features associated with human AITL. In this study we explored whether ROQUIN alterations could be an initial event in the human AITL oncogenic process. Using microarray and RT-PCR analyses, we investigated the levels of ROQUIN transcripts in TFH tumor cells purified from AITL (n = 8) and reactive tonsils (n = 12) and found similar levels of ROQUIN expression in both. Moreover, we also demonstrated that ROQUIN protein was expressed by AITL TFH (PD1+) cells. We then analysed ROQUIN coding sequence in 12 tumor cell-rich AITL samples and found no mutation in any of the samples. Finally, we analysed the expression of MiR101, a putative partner of ROQUIN involved in the modulation of ICOS expression and found similar levels of expression in tumor and reactive TFH. Altogether, this study shows that neither alteration of ROQUIN gene nor deregulation of miR101 expression is likely to be a frequent recurrent event in AITL.

Asunto(s)

Linfadenopatía Inmunoblástica/enzimología , Linfoma de Células T/enzimología , Ubiquitina-Proteína Ligasas/metabolismo , Células Cultivadas , Humanos , Linfadenopatía Inmunoblástica/genética , Linfadenopatía Inmunoblástica/metabolismo , Inmunohistoquímica , Linfoma de Células T/genética , Linfoma de Células T/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Ubiquitina-Proteína Ligasas/genética

RESUMEN

The ability of different synthetic cell penetrating peptides, as Antennapedia (wild and Phe(6) mutated penetratins), flock house virus, and integrin peptides to form complexes with a 25mer antisense oligonucleotide was compared and their conformation was determined by circular dichroism spectroscopy. The efficiency for oligonucleotide delivery into cells was measured using peptides labeled with a coumarin derivative showing blue fluorescence and the fluorescein-labeled antisense oligonucleotide showing green fluorescence. Fluorescence due to the excitation energy transfer confirmed the interaction of the antisense oligonucleotide and cell-penetrating peptides. The most efficient oligonucleotide delivery was found for penetratins. Comparison of the two types of penetratins shows that the wild-type penetratin proved to be more efficient than mutated penetratin. The paper also emphasizes that the attachment of a fluorescent label may have an effect on the conformation and flexibility of cell-penetrating peptides that must be taken into consideration when evaluating biological experiments.

Asunto(s)

Proteínas Portadoras/química , Proteínas Portadoras/farmacocinética , Portadores de Fármacos/química , Portadores de Fármacos/farmacocinética , Oligonucleótidos Antisentido/farmacocinética , Proteínas de Fusión Oncogénica/genética , Proteína Proto-Oncogénica c-fli-1/genética , Secuencia de Aminoácidos , Animales , Proteínas Portadoras/síntesis química , Proteínas Portadoras/genética , Línea Celular Transformada , Péptidos de Penetración Celular , Dicroismo Circular , Regulación hacia Abajo , Portadores de Fármacos/síntesis química , Técnicas de Transferencia de Gen , Ratones , Datos de Secuencia Molecular , Mutación , Células 3T3 NIH , Oligonucleótidos Antisentido/química , Oligonucleótidos Antisentido/genética , Conformación Proteica , Transporte de Proteínas , Proteína EWS de Unión a ARN