Manca, Paolo (2008) The Relative effectiveness of 2-Methoxyestradiol on undifferentiated and differentiated cells of glial and neuronal origin. Doctoral Thesis.
INTRODUCTION: 2-Methoxyestradiol (2ME) is a well-known metabolite of 17β–estradiol with a high cytoxic, antimitotic and apoptotic activity. AIMS: Find differences in the relative effectiveness of 2ME on undifferentiated fast-growing and differentiated slowgrowing cells of glial and neuronal origin. MATERIALS AND METHODS: Undifferentiated glial (clone C6)/neuronal(clone C1300) cells and tapsigargin-differentiated glial/db-cAMP differentiated neuronal cells were exposed at 2ME micromolar concentrations for 5 day, observed at the contrast phase microscope and the vitality test (Hoecst 33258/propidium iodide staining)/cell proliferation rate test (MTT) were carried out. The expression of total alpha tubulin, acetylated tubulin and tyrosinated tubulin was detected by western blot. RESULTS: Undifferentiated cells – 2ME was more effective in C1300 than in C6 cells; acetylated and mainly tyrosinated alpha tubulin downregulation was more intense in C1300 cells than in C6 cells. Differentiated cells – Cells were rather resistant to 2ME; total and tyrosinated alpha tubulin expression were slightly downregulated in neuronal cells whereas acetylated tubulin was upregulated both in glial/neuronal cells. CONCLUSIONS: 2ME is more effective in neuronal than in glial cells and in undifferentiated cells in comparison whit differentiated ones. 2ME toxicity may be related to the neuronal phenotypic and metabolic patterns, to the affinity for neuronal tubulin isoforms and to the high proliferation rate of undifferentiated cells. Moreover, intense downregulation of tyrosinated tubulin showed that 2ME alters selectively the microtubular dynamics.
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