The consensus sequence of FAMLF alternative splice variants is overexpressed in undifferentiated hematopoietic cells

The familial acute myeloid leukemia related factor gene (FAMLF) was previously identified from a familial AML subtractive cDNA library and shown to undergo alternative splicing. This study used real-time quantitative PCR to investigate the expression of the FAMLF alternative-splicing transcript consensus sequence (FAMLF-CS) in peripheral blood mononuclear cells (PBMCs) from 119 patients with de novo acute leukemia (AL) and 104 healthy controls, as well as in CD34+ cells from 12 AL patients and 10 healthy donors. A 429-bp fragment from a novel splicing variant of FAMLF was obtained, and a 363-bp consensus sequence was targeted to quantify total FAMLF expression. Kruskal-Wallis, Nemenyi, Spearman's correlation, and Mann-Whitney U-tests were used to analyze the data. FAMLF-CS expression in PBMCs from AL patients and CD34+ cells from AL patients and controls was significantly higher than in control PBMCs (P<0.0001). Moreover, FAMLF-CS expression in PBMCs from the AML group was positively correlated with red blood cell count (rs =0.317, P=0.006), hemoglobin levels (rs =0.210, P=0.049), and percentage of peripheral blood blasts (rs =0.256, P=0.027), but inversely correlated with hemoglobin levels in the control group (rs =–0.391, P<0.0001). AML patients with high CD34+ expression showed significantly higher FAMLF-CS expression than those with low CD34+ expression (P=0.041). Our results showed that FAMLF is highly expressed in both normal and malignant immature hematopoietic cells, but that expression is lower in normal mature PBMCs.

Análise da citodiferenciaçäo de linhagens celulares derivadas de adenoma pleomórfico e de mioepitelioma humanos de glândulas salivares: estudo comparativo dos elementos citoesqueléticos e da resposta dessas células à membrana basal reconstituída (Matrigel) / Cytodifferentiation analysis of cell lines derived from human pleomorphic adenoma and human myoepithelioma: a comparative study based on the expression of cytoskeletal components and on the response of these two cell lines to a reconstituted basement

Adenoma pleomórfico e mioepitelioma säo neoplasias de glândulas salivares que exibem aspectos clínicos e histológicos semelhantes. Para avaliar o estágio de diferenciaçäo das células de maior capacidade proliferativa dessas neoplasias, utilizamos linhagens celulares derivadas de adenoma pleomórfico (AP2) e de mioepitelioma (M1). Estudamos a expressäo de proteínas citoesqueléticas, os aspectos subcelulares e a resposta das linhagens à membrana basal reconstituída (Matrigel). Células AP2 mostraram imunomarcaçäo a vimentina e citoqueratina 14, enquanto que células M1 a vimentina, pan-queratina e actina de músculo liso. Estudo subcelular da linhagem AP2 revelou características de células pouco diferenciadas. Célula M1 mostraram aspectos subcelulares de fenótipo mioepitelial bem diferenciado, exibindo feixes de microfilamentos com adensamentos focais. Após uma semana envolvidas por Matrigel, células AP2 exibiram formaçäo de estruturas ductiformes e células M1 organizaram-se em cordöes celulares. Nossos resultados indicam que células AP2 exibem fenótipo epitelial glandular neoplásico pouco diferenciado, enquanto que células M1, fenótipo mioepitelial neoplásico bem diferenciado. Admitindo-se que o adenoma pleomórfico e o mioepitelioma resultem de proliferaçäo neoplástica de células do ducto intercalado de glândulas salivares, o adenoma pleomórfico originar-se-ia de células mais indiferenciadas e permissivas, enquanto que células já comprometidas com a diferenciaçäo no sentido mioepitelial dariam origem ao mioepitelioma