Navegando por Palavras-chave "In Situ Hybridization, Fluorescence"

Agora exibindo 1 - 1 de 1
  • Imagem de Miniatura
    Item
    Restrito
    Avaliação da Expressão da Vimentina em Leucemia Linfocítica Crônica e sua importância como fator de prognóstico
    (Universidade Federal de São Paulo (UNIFESP), 2010-02-24) Siufi, Grazziella Curado [UNIFESP]; Yamamoto, Mihoko [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Purpose: In this study our purpose was to evaluate the prognostic significance of the vimentin expression on CLL cells, from peripheral blood (PB) and bone marrow (BM) biopsy samples, correlating with already well known prognostic markers in CLL such as mutational status of IgVH genes, expression of CD38, Zap-70 protein, and VEGFR, molecular cytogenetics (FISH) results and with Binet’s clinical stage and progression free survival (PFS) Methods: Sixty six patients with CLL from Hospital São Paulo-UNIFESP were studied. The CLL diagnosis was established by leukemic cells immunophenotyping using score system (Moreau et al). PB cells were collected at admission and evaluated by flow cytometry for CD38 (through positive cells percentage and mean fluorescence intensity –MFI- methods) and Zap-70, VEGFR and Vimentin expressions. For the last ones (Vimentin and VEGFR) mononuclear cells separated by Ficoll Hypaque gradient were used. The Vimentin expression was evaluated in fresh and/or cryopreserved blood samples and also in BM biopsy specimens obtained at the time of diagnosis, by immunohistochemistry.The BM infiltration pattern was also evaluated. The IgVH genes mutational status was evaluated by PCR techniques followed by sequencing. Results: Vimentin expression in both circulating and marrow cells was associated with VEGFR positive expression, unmutated IgVH genes, diffuse bone marrow infiltration and unfavourable cytogenetics abnormalities. Association of CD38 expression was observed only in BM. Advanced clinical stage, age>68 years, positive expression for CD38 and VEGFR, unmutated IgVH genes, unfavourable cytogenetics abnormalities, vimentin expression in PB and BM cells and diffuse marrow infiltration pattern were associated with short PFS. The impact of these prognostic markers was also observed in Binet A patients. In multivariate analysis, the Binet’s stages B+C (p=0,008), strong unimodal and bimodal IMF pattern of CD38 (p=0,03), diffuse marrow infiltration (p=0,02) and vimentin expression on BM cells by immunohistochemistry (p=0,002) were identified as independent risk factor for reduced PFS. Comparison of Vimentin expression in PB cells by flow cytometry and in BM cells by immunohistochemistry from the same patients, showed concordant results. When vimentin expression in fresh and thawed samples from the same patients was compared, concordant results were also observed. Conclusions: Vimentin expression in CLL cells from both PB and BM showed high impact as prognostic marker, predicting poor PFS, even in Binet A patients. Both flow cytometry and immunohistochemistry are relatively easy techniques and feasible in most flow cytometric or pathological laboratories, requiring the usual routine equipments. Our results were comparable to IgVH mutational status suggesting the usefulness of Vimentin as prognostic marker in CLL patients. In addition its evaluation might be useful for the better understanding of pathofisiology of CLL and also, for the therapeutic interventions, such as the use of EGCG (epigallocatechin galate – green tea).